Substituted piperidines

ABSTRACT

The invention relates to novel substituted piperidines, to processes for preparation thereof, to the use thereof for treatment and/or prophylaxis of diseases and to the use thereof for production of medicaments for treatment and/or prophylaxis of diseases, especially of cardiovascular diseases and tumour diseases.

The invention relates to novel substituted piperidines, to processes forpreparation thereof, to the use thereof for treatment and/or prophylaxisof diseases and to the use thereof for production of medicaments fortreatment and/or prophylaxis of diseases, especially of cardiovasculardiseases and tumour diseases.

Thrombocytes (blood platelets) are a significant factor both inphysiological haemostasis and in thromboembolic disorders. In thearterial system in particular, platelets are of central importance inthe complex interaction between blood components and the wall of thevessel. Unwanted platelet activation may, as a result of formation ofplatelet-rich thrombi, lead to thromboembolic disorders and thromboticcomplications with life-threatening conditions.

One of the most potent platelet activators is the blood coagulationprotease thrombin, which is formed at injured blood vessel walls andwhich, in addition to fibrin formation, leads to the activation ofplatelets, endothelial cells and mesenchymal cells (Vu T K H, Hung D T,Wheaton V I, Coughlin S R, Cell 1991, 64, 1057-1068). In platelets invitro and in animal models, thrombin inhibitors inhibit plateletaggregation and the formation of platelet-rich thrombi. In man, arterialthromboses can be prevented or treated successfully with inhibitors ofplatelet function and thrombin inhibitors (Bhatt D L, Topol E J, Nat.Rev. Drug Discov. 2003, 2, 15-28). Accordingly, there is a highprobability that antagonists of thrombin action on blood plateletsreduce the formation of thrombi and the occurrence of clinical sequelaesuch as myocardial infarction and stroke. Other cellular effects ofthrombin, for example on endothelial and smooth muscle cells of vessels,leukocytes and fibroblasts, are possibly responsible for inflammatoryand proliferative disorders.

At least some of the cellular effects of thrombin are mediated via afamily of G-protein-coupled receptors (Protease Activated Receptors,PARs), the prototype of which is the PAR-1 receptor. PAR-1 is activatedby bindung of thrombin and proteolytic cleavage of its extracellularN-terminus. The proteolysis exposes a new N-terminus having the aminoacid sequence SFLLRN . . . , which, as agonist (“tethered ligand”),leads to intramolecular receptor activation and transmission ofintracellular signals. Peptides derived from the tethered ligandsequence can be used as agonists of the receptor and, on platelets, leadto activation and aggregation. Other proteases are likewise capable ofactivating PAR-1; these proteases include, for example, plasmin, factorVIIa, factor Xa, trypsin, activated protein C (aPC), tryptase, cathepsinG, proteinase 3, granzyme A, elastase and matrix metalloprotease 1(MMP-1).

In contrast to the inhibition of protease activity of thrombin withdirect thrombin inhibitors, blockade of PAR-1 should result in aninhibition of platelet activation without reduction in the coagulabilityof the blood (anticoagulation).

Antibodies and other selective PAR-1 antagonists inhibit thethrombin-induced aggregation of platelets in vitro at low to mediumthrombin concentrations (Kahn M L, Nakanishi-Matsui M, Shapiro M J,Ishihara H, Coughlin S R, J. Clin. Invest. 1999, 103, 879-887). Afurther thrombin receptor with possible significance for thepathophysiology of thrombotic processes, PAR-4, was identified on humanand animal platelets. In experimental thromboses in animals having a PARexpression pattern comparable to humans, PAR-1 antagonists reduce theformation of platelet-rich thrombi (Derian C K, Damiano B P, Addo M F,Darrow A L, D'Andrea M R, Nedelman M, Zhang H-C, Maryanoff B E,Andrade-Gordon P, J. Pharmacol. Exp. Ther. 2003, 304, 855-861).

In the last few years, a large number of substances have been examinedfor their platelet function-inhibiting action; but only a few plateletfunction inhibitors have been found to be useful in practice. There istherefore a need for pharmaceuticals which specifically inhibit anincreased platelet reaction without significantly increasing the risk ofbleeding, thus reducing the risk of thromboembolic complications.

Effects of thrombin which are mediated via the PAR-1 receptor influencethe progression of the disease during and after coronary artery bypassgraft (CABG) and other operations, and in particular operations withextracorporeal circulation (for example heart-lung machine). During thecourse of the operation, there may be bleeding complications owing topre- or intraoperative medication with coagulation-inhibiting and/orplatelet-inhibiting substances. For this reason, for example, medicationwith clopidogrel has to be interrupted several days prior to a CABG.Moreover, as mentioned, disseminated intravascular coagulation orconsumption coagulopathy (DIC) may develop (for example owing to theextended contact between blood and synthetic surfaces duringextracorporeal circulation or blood transfusions), which in turn canlead to bleeding complications. At a later stage, there is frequentlyrestenosis of the venous or arterial bypasses grafted (which may evenresult in occlusion) owing to thrombosis, intimafibrosis,arteriosclerosis, angina pectoris, myocardial infarction, heart failure,arrhythmias, transitory ischaemic attack (TIA) and/or stroke.

In man, the PAR-1 receptor is also expressed in other cells including,for example, endothelial cells, smooth muscle cells and tumour cells.Malignant tumour disorders (cancer) have a high incidence and aregenerally associated with high mortality. Current treatments achievefull remission in only a fraction of patients and are typicallyassociated with severe side effects. There is therefore a high demandfor more effective and safer therapies. The PAR-1 receptor contributesto cancer generation, growth, invasiveness and metastasis. Moreover,PAR-1 expressed on endothelial cells mediates signals resulting invascular growth (“angiogenesis”), a process which is vital for enablingtumour growth beyond about 1 mm³ Angiogenesis also contributes to thegenesis or worsening of other disorders including, for example,haematopoetic cancer disorders, macular degeneration, which leads toblindness, and diabetic retinopathy, inflammatory disorders, such asrheumatoid arthritis and colitis.

Sepsis (or septicaemia) is a common disorder with high mortality.Initial symptoms of sepsis are typically unspecific (for example fever,reduced general state of health); however, during further progressionthere may be generalized activation of the coagulation system(“disseminated intravascular coagulation” or “consumption coagulopathy”(DIC)) with the formation of microthrombi in various organs andsecondary bleeding complications. DIC may also occur independently ofsepsis, for example during operations or associated with tumourdisorders.

Treatment of sepsis consists firstly in the rigorous elimination of theinfectious cause, for example by operative focal reconstruction andantibiosis. Secondly, it consists in temporary intensive medical supportof the affected organ systems. Treatments of the different stages ofthis disease have been described, for example, in the followingpublication (Dellinger et al., Crit. Care Med. 2004, 32, 858-873). Thereare no proven effective treatments for DIC.

It is therefore an object of the present invention to provide novelPAR-1 antagonists for the treatment of disorders, for examplecardiovascular disorders and thromboembolic disorders, and also tumourdisorders in humans and animals.

WO 2006/012226, WO 2006/020598, WO 2007/038138, WO 2007/130898, WO2007/101270 and US 2006/0004049 describe structurally similarpiperidines as 11-β HSD1 inhibitors for treatment of, inter alia,diabetes, thromboembolic disorders and stroke.

The invention provides compounds of the formula

in which

-   R¹ is trifluoromethyl, 1,1-difluoroethyl, 2,2-difluoroethyl,    2,2,2-trifluoroethyl, difluoromethoxy, trifluoromethoxy or ethyl,-   R² is 2-hydroxyeth-1-yl, 2-methoxyeth-1-yl, 2-ethoxyeth-1-yl,    cyclopropyl or 1-methoxycycloprop-1-yl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Inventive compounds are the compounds of the formula (I) and theirsalts, solvates and solvates of the salts; the compounds, encompassed byformula (I), of the formulae mentioned below and their salts, solvatesand solvates of the salts, and the compounds encompassed by the formula(I), cited below as working examples, and their salts, solvates andsolvates of the salts if the compounds, encompassed by the formula (I),cited below are not already salts, solvates and solvates of the salts.

Depending on their structure, the inventive compounds may exist instereoisomeric forms (enantiomers, diastereomers). Accordingly, theinvention encompasses the enantiomers or diastereomers and theirrespective mixtures. From such mixtures of enantiomers and/ordiastereomers, it is possible to isolate the stereoisomerically uniformconstituents in a known manner.

If the inventive compounds can occur in tautomeric forms, the presentinvention encompasses all tautomeric forms.

In the context of the present invention, preferred salts arephysiologically acceptable salts of the inventive compounds. Alsoencompassed, however, are salts which for their part are not suitablefor pharmaceutical applications, but which can be used, for example, forisolating or purifying the inventive compounds.

Physiologically acceptable salts of the inventive compounds include acidaddition salts of mineral acids, carboxylic acids and sulphonic acids,for example salts of hydrochloric acid, hydrobromic acid, sulphuricacid, phosphoric acid, methanesulphonic acid, ethanesulphonic acid,toluenesulphonic acid, benzenesulphonic acid, naphthalenedisulphonicacid, acetic acid, trifluoroacetic acid, propionic acid, lactic acid,tartaric acid, malic acid, citric acid, fumaric acid, maleic acid andbenzoic acid.

Physiologically acceptable salts of the inventive compounds also includesalts of customary bases, such as, by way of example and withpreference, alkali metal salts (for example sodium salts and potassiumsalts), alkaline earth metal salts (for example calcium salts andmagnesium salts) and ammonium salts, derived from ammonia or organicamines having 1 to 16 carbon atoms, such as, by way of example and withpreference, ethylamine, diethylamine, triethylamine,ethyldiisopropylamine, monoethanolamine, diethanolamine,triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine,dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine,N-methylpiperidine and choline.

In the context of the invention, solvates refer to those forms of theinventive compounds which, in solid or liquid state, form a complex bycoordination with solvent molecules. Hydrates are a specific form of thesolvates in which the coordination is with water.

Moreover, the present invention also encompasses prodrugs of theinventive compounds. The term “prodrugs” includes compounds which fortheir part may be biologically active or inactive but which, during thetime they spend in the body, are converted to inventive compounds (forexample metabolically or hydrolytically).

In the formula of the group which may be R³, the end point of the linemarked by * does not represent a carbon atom or a CH₂ group, but is partof the bond to the atom to which R³ is attached.

Preference is given to compounds of the formula (I) in which

-   R¹ is trifluoromethyl, 1,1-difluoroethyl, 2,2-difluoroethyl,    2,2,2-trifluoroethyl, difluoromethoxy, trifluoromethoxy or ethyl,-   R² is 2-methoxyeth-1-yl, cyclopropyl or 1-methoxycycloprop-1-yl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which

-   R¹ is trifluoromethyl, 2,2,2-trifluoroethyl, trifluoromethoxy or    ethyl,-   R² is 2-methoxyeth-1-yl, cyclopropyl or 1-methoxycycloprop-1-yl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which

-   R¹ is trifluoromethyl, 2,2,2-trifluoroethyl or trifluoromethoxy,-   R² is cyclopropyl or 1-methoxycycloprop-1-yl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which

-   R¹ is trifluoromethyl, trifluoromethoxy or ethyl,-   R² für 2-hydroxyeth-1-yl, 2-methoxyeth-1-yl, 2-ethoxyeth-1-yl or    cyclopropyl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which

-   R¹ is trifluoromethyl or ethyl,-   R² is 2-hydroxyeth-1-yl, 2-methoxyeth-1-yl or cyclopropyl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which

-   R¹ is trifluoromethyl or ethyl,-   R² is 2-methoxyeth-1-yl,-   R³ is a group of the formula

-   -   where        * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which

-   R¹ is trifluoromethyl or ethyl,-   R² is 2-methoxyeth-1-yl,-   R³ is a group of the formula

-   -   where        * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is given to compounds of the formula (I) in which

-   R¹ is trifluoromethoxy,-   R² is 2-methoxyeth-1-yl or cyclopropyl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is given to compounds of the formula (I) in which

-   R¹ is trifluoromethoxy,-   R² is cyclopropyl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is given to compounds of the formula (I) in which

-   R¹ is trifluoromethoxy,-   R² is 2-methoxyeth-1-yl or cyclopropyl,-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group,        and their salts, their solvates and the solvates of their salts.

Preference is also given to compounds of the formula (I) in which thephenyl substituent and 1,2,4-oxadiazol-5-yl substituent, which arebonded to the piperidine ring, are in cis positions to one another.

Preference is also given to compounds of the formula (I) in which thecarbon atom to which the phenyl substituent is bonded has Sconfiguration and the carbon atom to which the 1,2,4-oxadiazol-5-ylsubstituent is bonded likewise has S configuration.

Preference is also given to compounds of the formula (I) in which R¹ istrifluoromethyl.

Preference is also given to compounds of the formula (I) in which R¹ istrifluoromethoxy.

Preference is also given to compounds of the formula (I) in which R¹ isethyl.

Preference is also given to compounds of the formula (I) in which R² is2-methoxyeth-1-yl.

Preference is also given to compounds of the formula (I) in which R² iscyclopropyl.

Preference is also given to compounds of the formula (I) in which R² is1-methoxycycloprop-1-yl.

Preference is also given to compounds of the formula (I) in which

R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group.        Preference is also given to compounds of the formula (I) in        which

-   R³ is a group of the formula

-   -   where    -   * is the point of attachment to the carbonyl group.

The individual radical definitions specified in the respectivecombinations or preferred combinations of radicals are, independently ofthe respective combinations of the radicals specified, also replaced asdesired by radical definitions of other combinations.

Very particular preference is given to combinations of two or more ofthe preferred ranges mentioned above.

The invention further provides a process for preparing the compounds ofthe formula (I), or their salts, their solvates or the solvates of theirsalts, wherein

[A] compounds of the formula

in whichR¹ and R² are each as defined aboveare reacted with compounds of the formula

in whichR³ is as defined above andX¹ is halogen, preferably bromine or chlorine, or hydroxyl or4-nitrophenoxy,or[B] compounds of the formula (II) are reacted in the first stage with4-nitrophenyl chloroformate and in the second stage with compounds ofthe formula

R³—H  (IV)

in whichR³ is as defined above,or[C] compounds of the formula

in whichR¹ and R³ are each as defined aboveare reacted with compounds of the formula

in whichR² is as defined above,or[D] compounds of the formula

in whichR¹ and R² are each as defined aboveare reacted with 0.8 to 1.1 equivalents of meta-chloroperbenzoic acid togive compounds of the formula

in whichR¹ and R² are each as defined aboveor[E] compounds of the formula (Ia) are reacted with 2.0 to 3.0equivalents of meta-chloroperbenzoic acid to give compounds of theformula

in whichR¹ and R² are each as defined above.

The compounds of the formulae (Ia), (Ib) and (Ic) are a subset of thecompounds of the formula (I).

When X¹ is halogen, the reaction according to method [A] is generallyeffected in inert solvents, optionally in the presence of a base,preferably in a temperature range of −30° C. to 50° C. at standardpressure.

Inert solvents are, for example, tetrahydrofuran, methylene chloride,pyridine, dioxane or dimethylformamide, preference being given tomethylene chloride.

Bases are, for example, triethylamine, diisopropylethylamine orN-methylmorpholine, preference being given to triethylamine ordiisopropylethylamine

When X¹ is hydroxyl, the reaction according to method [A] is generallyeffected in inert solvents, in the presence of a dehydrating reagent,optionally in the presence of a base, preferably in a temperature rangeof −30° C. to 50° C. at standard pressure.

Inert solvents are, for example, halohydrocarbons such asdichloromethane or trichloromethane, hydrocarbons such as benzene,nitromethane, dioxane, dimethylformamide or acetonitrile. It is equallypossible to use mixtures of the solvents. Particular preference is givento dichloromethane or dimethylformamide.

Suitable dehydrating reagents in this context are, for example,carbodiimides, for example N,N′-diethyl-, N,N′-dipropyl-,N,N′-diisopropyl-, N,N′-dicyclohexylcarbodiimide,N-(3-dimethylamino-isopropyl)-N′-ethylcarbodiimide hydrochloride (EDC),N-cyclohexylcarbodiimide-N′-propyloxymethylpolystyrene(PS-carbodiimide), or carbonyl compounds such as carbonyldiimidazole, or1,2-oxazolium compounds such as 2-ethyl-5-phenyl-1,2-oxazolium3-sulphate or 2-tert-butyl-5-methylisoxazolium perchlorate, or acylaminocompounds such as 2-ethoxy-1-ethoxy-carbonyl-1,2-dihydroquinoline, orpropanephosphonic anhydride, or isobutyl chloroformate, orbis-(2-oxo-3-oxazolidinyl)phosphoryl chloride orbenzotriazolyloxytri(dimethylamino)phosphonium hexafluorophosphate, orO-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate(HBTU), 2-(2-oxo-1-(2H)-pyridyl)-1,1,3,3-tetramethyluroniumtetrafluoroborate (TPTU) orO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU), or 1-hydroxy-benzotriazole (HOBt), orbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP), or N-hydroxysuccinimide, or mixtures of these, with bases.

Bases are, for example, alkali metal carbonates, for example sodiumcarbonate or potassium carbonate, or sodium hydrogencarbonate orpotassium hydrogencarbonate, or organic bases such as trialkylamines,for example triethylamine, N-methylmorpholine, N-methylpiperidine,4-dimethylaminopyridine or diisopropylethylamine.

Preferably, the condensation is effected with HATU or with EDC in thepresence of HOBt.

When X¹ is 4-nitrophenoxy, the reaction according to method [A] isgenerally effected in inert solvents, optionally in the presence of abase, optionally in a microwave, preferably in a temperature range of50° C. to 200° C. at standard pressure to 5 bar.

Inert solvents are, for example, N-methylpyrrolidone, dioxane ordimethylformamide, preference being given to N-methylpyrrolidone.

Bases are, for example, triethylamine, diisopropylethylamine orN-methylmorpholine, preference being given to triethylamine ordiisopropylethylamine

The compounds of the formula (III) are known or can be synthesized byknown processes from the appropriate starting compounds.

The reaction of the first stage according to method [B] is generallyeffected in inert solvents, in the presence of a base, preferably in atemperature range of 0° C. to 50° C. at standard pressure.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane, carbon tetrachloride or 1,2-dichloroethane,preference being given to methylene chloride.

Bases are, for example, organic bases such as trialkylamines, forexample triethylamine, N-methyl-morpholine, N-methylpiperidine,4-dimethylaminopyridine or diisopropylethylamine, preference being givento triethylamine.

The reaction of the second stage according to method [B] is generallyeffected in inert solvents, in the presence of a base, optionally in amicrowave, preferably in a temperature range of 50° C. to 200° C. atstandard pressure to 5 bar.

Inert solvents are, for example, dimethyl sulphoxide, dimethylformamideor N-methylpyrrolidone, preference being given to dimethylformamide.

Bases are, for example, alkali metal carbonates, for example sodiumcarbonate or potassium carbonate, preference being given to potassiumcarbonate.

The compounds of the formula (IV) are known or can be synthesized byknown processes from the appropriate starting compounds.

The reaction according to method [C] is generally effected in inertsolvents, in the presence of a dehydrating reagent, optionally in thepresence of a base, preferably in a temperature range from roomtemperature up to reflux of the solvents at standard pressure.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane or 1,2-dichloroethane, ethers such asdioxane, tetrahydrofuran or 1,2-dimethoxyethane, or other solvents suchas acetone, dimethylformamide, dimethylacetamide, 2-butanone oracetonitrile. It is equally possible to use mixtures of the solvents.Preference is given to dimethylformamide or a mixture of dioxane anddimethylformamide.

Suitable dehydrating reagents in this context are, for example,carbodiimides, for example N,N′-diethyl-, N,N′-dipropyl-,N,N′-diisopropyl-, N,N′-dicyclohexylcarbodiimide,N-(3-dimethylamino-isopropyl)-N′-ethylcarbodiimide hydrochloride (EDC),N-cyclohexylcarbodiimide-N′-propyloxymethylpolystyrene(PS-carbodiimide), or carbonyl compounds such as carbonyldiimida-zole,or 1,2-oxazolium compounds such as 2-ethyl-5-phenyl-1,2-oxazolium3-sulphate or 2-tert-butyl-5-methylisoxazolium perchlorate, or acylaminocompounds such as 2-ethoxy-1-ethoxy-carbonyl-1,2-dihydroquinoline, orpropanephosphonic anhydride, or isobutyl chloroformate, orbis-(2-oxo-3-oxazolidinyl)phosphoryl chloride orbenzotriazolyloxytri(dimethylamino)phosphonium hexafluorophosphate, orO-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate(HBTU), 2-(2-oxo-1-(2H)-pyridyl)-1,1,3,3-tetramethyluroniumtetrafluoroborate (TPTU) orO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU), or 1-hydroxy-benzotriazole (HOBt), orbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP), or benzotriazol-1-yloxytris(pyrrolidino)phosphoniumhexafluorophosphate (PYBOP), or N-hydroxysuccinimide, or mixtures ofthese with bases.

Bases are, for example, alkali metal carbonates, for example sodiumcarbonate or potassium carbonate, or sodium hydrogencarbonate orpotassium hydrogencarbonate, or organic bases such as trialkylamines,for example triethylamine, N-methylmorpholine, N-methylpiperidine,4-dimethylaminopyridine or diisopropylethylamine, preference being givento diisopropylethylamine.

Preferably, the condensation is carried out with HATU in the presence ofdiisopropylethylamine or alternatively only with carbonyldiimidazole.

The compounds of the formula (VI) are known or can be synthesized byknown processes from the appropriate starting compounds.

The reaction according to method [D] is generally performed in inertsolvents, preferably in a temperature range from room temperature up toreflux of the solvents at standard pressure.

meta-Chloroperbenzoic acid is preferably used in an amount of 0.9 to 1.0equivalent.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane or 1,2-dichloroethane. Preference is given tomethylene chloride.

The reaction according to method [E] is generally effected in inertsolvents, preferably in a temperature range from room temperature up toreflux of the solvents at standard pressure.

meta-Chloroperbenzoic acid is preferably used in an amount of 2.3 to 2.6equivalents, more preferably in an amount of 2.5 equivalents.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane or 1,2-dichloroethane. Preference is given tomethylene chloride.

The compounds of the formula (II) are known or can be prepared byreacting compounds of the formula

in whichR¹ is as defined abovein the first stage with compounds of the formula (VI) and in the secondstage with an acid.

The first stage reaction is effected as described for process [C]

The second stage reaction is generally effected in inert solvents,preferably in a temperature range from room temperature to 60° C. atstandard pressure.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane, carbon tetrachloride or 1,2-dichloroethane,or ethers such as tetrahydrofuran or dioxane, preference being given tomethylene chloride.

Bases are, for example, trifluoroacetic acid or hydrogen chloride indioxane, preference being given to trifluoroacetic acid.

The compounds of the formula (VII) are known or can be prepared byreacting compounds of the formula

in whichR¹ is as defined above andR⁴ is methyl or ethyl,in the first stage with di-tert-butyl dicarboxylate andin the second stage with a base.

The first stage reaction is generally effected in inert solvents, in thepresence of a base, preferably in a temperature range from roomtemperature to 50° C. at standard pressure.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane, carbon tetrachloride or 1,2-dichloroethane,preference being given to methylene chloride.

Bases are, for example, triethylamine, diisopropylethylamine orN-methylmorpholine, preference being given to triethylamine ordiisopropylethylamine

The second stage reaction is generally effected in inert solvents, inthe presence of a base, preferably in a temperature range from roomtemperature up to reflux of the solvents at standard pressure.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane, tetrachloromethane or 1,2-dichloroethane,alcohols such as methanol or ethanol, ethers such as diethyl ether,methyl tert-butyl ether, 1,2-dimethoxyethane, dioxane ortetrahydrofuran, or other solvents such as dimethylformamide,dimethylacetamide, acetonitrile or pyridine, or mixtures of solvents, ormixtures of solvent with water, preference being given to methanol ormethanol with one equivalent of water, or a mixture of tetrahydrofuranand water.

Bases are, for example, alkali metal hydroxides such as sodium, lithiumor potassium hydroxide, or alkali metal carbonates such as caesiumcarbonate, sodium or potassium carbonate, or alkoxides such as potassiumor sodium tert-butoxide, preference being given to lithium hydroxide orpotassium tert-butoxide.

The compounds of the formula (VIII) are known or can be prepared byhydrogenating compounds of the formula

in whichR¹ and R⁴ are each as defined above.

The hydrogenation is generally effected with a reducing agent in inertsolvents, optionally with addition of acid such as mineral acids andcarboxylic acids, preferably acetic acid, preferably in a temperaturerange from room temperature up to reflux of the solvents and in apressure range from standard pressure to 100 bar, preferably at standardpressure or at 50-80 bar.

A preferred reducing agent is hydrogen with palladium on activatedcarbon, with rhodium on activated carbon, with ruthenium on activatedcarbon or mixed catalysts thereof, or hydrogen with palladium on aluminaor with rhodium on alumina, or hydrogen with palladium on activatedcarbon and platinum(IV) oxide, preference being given to hydrogen withpalladium on activated carbon or with rhodium on activated carbon orhydrogen with palladium on activated carbon and platinum(IV) oxide. Itis also possible to hydrogenate under pressure with hydrogen andplatinum(IV) oxide alone.

Inert solvents are, for example, alcohols such as methanol, ethanol,n-propanol, isopropanol, n-butanol or tert-butanol, or concentratedacetic acid or methanol with addition of concentrated hydrochloric acid,preference being given to methanol or ethanol or concentrated aceticacid or methanol with addition of concentrated hydrochloric acid.

The compounds of the formula (IX) are known or can be prepared byreacting compounds of the formula

in whichR⁴ is as defined abovewith compounds of the formula

in whichR¹ is as defined above.

The reaction is generally effected in inert solvents, in the presence ofa catalyst, if appropriate in the presence of an additional reagent,preferably in a temperature range from room temperature up to reflux ofthe solvent at standard pressure.

Inert solvents are, for example, ethers such as dioxane, tetrahydrofuranor 1,2-dimethoxyethane, hydrocarbons such as benzene, xylene or toluene,or other solvents such as nitrobenzene, dimethylformamide,dimethylacetamide, dimethyl sulphoxide or N-methylpyrrolidone; ifappropriate, some water is added to these solvents. Preference is givento toluene with water or to a mixture of 1,2-dimethoxyethane,dimethylformamide and water.

Catalysts are, for example, palladium catalysts customary for Suzukireaction conditions, preference being given to catalysts such asdichlorobis(triphenylphosphine)palladium,tetrakistriphenylphosphinepalladium(0), palladium(II) acetate orbis(diphenylphosphine-ferrocenyl)palladium(II) chloride, for example.

Additional reagents are, for example, potassium acetate, caesium,potassium or sodium carbonate, barium hydroxide, potassiumtert-butoxide, caesium fluoride, potassium fluoride or potassiumphosphate, or mixtures thereof, preference being given to potassiumfluoride or sodium carbonate, or a mixture of potassium fluoride andpotassium carbonate.

The compounds of the formulae (X), (XI) and (XIII) are known or can besynthesized by known processes from the appropriate starting compounds.

The compounds of the formula (V) are known or can be prepared byreacting compounds of the formula

in whichR¹ and R³ are each as defined above andR⁴ is methyl or ethyl,with a base.

The reaction is generally effected in inert solvents, in the presence ofa base, preferably in a temperature range from room temperature up toreflux of the solvents at standard pressure.

Inert solvents are, for example, halohydrocarbons such as methylenechloride, trichloromethane, tetrachloromethane or 1,2-dichloroethane,alcohols such as methanol or ethanol, ethers such as diethyl ether,methyl tert-butyl ether, 1,2-dimethoxyethane, dioxane ortetrahydrofuran, or other solvents such as dimethylformamide,dimethylacetamide, acetonitrile or pyridine, or mixtures of solvents, ormixtures of solvent with water, preference being given to methanol ormethanol with one equivalent of water, or a mixture of tetrahydrofuranand water.

Bases are, for example, alkali metal hydroxides such as sodium, lithiumor potassium hydroxide, or alkali metal carbonates such as caesiumcarbonate, sodium or potassium carbonate, or alkoxides such as potassiumor sodium tert-butoxide, preference being given to lithium hydroxide orpotassium tert-butoxide.

The compounds of the formula (XII) are known or can be prepared byreacting compounds of the formula (VIII) with compounds of the formula(III).

The reaction is effected as described for method [A].

In an alternative method, the compounds of the formula (XII) can beprepared by reacting compounds of the formula (VIII) in the first stagewith 4-nitrophenyl chloroformate and in the second stage with compoundsof the formula (IV).

The reaction is effected as described for method [B].

The preparation of the compounds of the formula (I) can be illustratedby the synthesis scheme below.

The inventive compounds have an unforeseeable, valuable spectrum ofpharmacological and pharmacokinetic activity. They are selectiveantagonists of the PAR-1 receptor acting in particular as plateletaggregation inhibitors, as inhibitors of endothelial cell activation, asinhibitors of smooth muscle cell proliferation and as inhibitors oftumour growth. For some of the diseases mentioned, for examplecardiovascular diseases with high thromboembolic risk, permanentprotection by PAR-1 antagonism with simultaneously simple management ofmedication is of great significance. The PAR-1 antagonists of thepresent invention exhibit long-lasting action after single oraladministration, i.e. an action which lasts at least 16 hours.

Accordingly, they are suitable for use as medicaments for treatmentand/or prophylaxis of diseases in man and animals.

The present invention further provides for the use of the inventivecompounds for treatment and/or prophylaxis of disorders, preferably ofthromboembolic disorders and/or thromboembolic complications.

“Thromboembolic disorders” in the context of the present inventioninclude in particular disorders such as ST-segment elevation myocardialinfarction (STEMI) and non-ST-segment elevation myocardial infarction(non-STEMI), stable angina pectoris, unstable angina pectoris,reocclusions and restenoses after coronary interventions such asangioplasty, stent implantations or aortocoronary bypass, peripheralarterial occlusion diseases, pulmonary embolisms, deep venous thrombosesand renal vein thromboses, transitory ischaemic attacks and alsothrombotic and thromboembolic stroke.

The substances are therefore also suitable for prevention and treatmentof cardiogenic thrombo-embolisms, for example brain ischaemias, strokeand systemic thromboembolisms and ischaemias, in patients with acute,intermittent or persistent cardiac arrhythmias, for example atrialfibrillation, and those undergoing cardioversion, and also in patientswith heart valve disorders or with intravasal objects, for exampleartificial heart valves, catheters, intraaortic balloon counterpulsationand pacemaker probes.

Thromboembolic complications are also encountered in connection withmicroangiopathic haemolytic anaemias, extracorporeal circulation, forexample haemodialysis, haemofiltration, ventricular assist devices andartificial hearts, and also heart valve prostheses.

Moreover, the inventive compounds are also used for influencing woundhealing, for the prophylaxis and/or treatment of atheroscleroticvascular disorders and inflammatory disorders such as rheumaticdisorders of the locomotive system, coronary heart diseases, of heartfailure, of hypertension, of inflammatory disorders, for example asthma,COPD, inflammatory pulmonary disorders, glomerulonephritis andinflammatory intestinal disorders, and additionally also for theprophylaxis and/or treatment of Alzheimer's disease, autoimmunedisorders, Crohn's disease and ulcerative colitis.

Moreover, the inventive compounds can be used for inhibiting tumourgrowth and the formation of metastases, for microangiopathies,age-related macular degeneration, diabetic retinopathy, diabeticnephropathy and other microvascular disorders, and also for theprevention and treatment of thromboembolic complications, for examplevenous thromboembolisms, for tumour patients, in particular thoseundergoing major surgical interventions or chemo- or radiotherapy.

The inventive compounds are additionally suitable for treatment ofcancer. Cancers include: carcinomas (including breast cancer,hepatocellular carcinomas, lung cancer, colorectal cancer, cancer of thecolon and melanomas), lymphomas (for example non-Hodgkin's lymphomas andmycosis fungoides), leukaemias, sarcomas, mesotheliomas, brain cancer(for example gliomas), germinomas (for example testicular cancer andovarian cancer), choriocarcinomas, renal cancer, cancer of the pancreas,thyroid cancer, head and neck cancer, endometrial cancer, cervicalcancer, bladder cancer, stomach cancer and multiple myeloma.

Moreover, PAR-1 expressed on endothelial cells mediates signalsresulting in vascular growth (“angiogenesis”), a process which is vitalfor enabling tumour growth beyond about 1 mm³.

Induction of angiogenesis is also relevant for other disorders; theseinclude disorders of the rheumatic type (for example rheumatoidarthritis), pulmonary disorders (for example pulmonary fibrosis,pulmonary hypertension, in particular pulmonary arterial hypertension,disorders characterized by pulmonary occlusion), arteriosclerosis,plaque rupture, diabetic retinopathy and wet macular degeneration.

In addition, the inventive compounds are suitable for treatment ofsepsis. Sepsis (or septicaemia) is a common disorder with highmortality. Initial symptoms of sepsis are typically unspecific (forexample fever, reduced general state of health); however, during furtherprogression there may be a general activation of the coagulation system(“disseminated intravascular coagulation” or “consumption coagulopathy”;referred to hereinafter as “DIC”) with the formation of microthrombi invarious organs and secondary bleeding complications. Moreover, there maybe endothelial damage with increased permeability of the vessels anddiffusion of fluid and proteins into the extravasal space. Later, theremay be organ dysfunction or organ failure (for example kidney failure,liver failure, respiratory failure, deficits of the central nervoussystem and heart/circulatory failure) and even multiple organ failure.In principle, this may affect any organ; the most frequently encounteredorgan dysfunctions and organ failures are those of the lung, the kidney,the cardiovascular system, the coagulation system, the central nervoussystem, the endocrine glands and the liver. Sepsis may be associatedwith an “acute respiratory distress syndrome” (referred to hereinafteras ARDS). ARDS may also occur independently of sepsis. “Septic shock”refers to the occurrence of hypotension, which requires treatment, andpromotes further organ damage and is associated with a worsening of theprognosis.

Pathogens may be bacteria (gram-negative and gram-positive), fungi,viruses and/or eukaryotes. The site of entry or primary infection may bepneumonia, an infection of the urinary tract or peritonitis, forexample. The infection may, but need not necessarily, be associated withbacteriaemia.

Sepsis is defined as the presence of an infection and a “systemicinflammatory response syndrome” (referred to hereinafter as “SIRS”).SIRS occurs during infections, but also during other states such asinjuries, burns, shock, operations, ischaemia, pancreatitis, reanimationor tumours. The definition of the ACCP/SCCM Consensus ConferenceCommittee of 1992 (Crit. Care Med. 1992, 20, 864-874) describes thesymptoms required for the diagnosis “SIRS” and measurement parameters(inter alia a change in body temperature, increased heart rate,breathing difficulties and altered blood profile). The later (2001)SCCM/ESICM/ACCP/ATS/SIS International Sepsis Definitions Conferenceessentially maintained the criteria, but fine-tuned details (Levy etal., Crit. Care Med. 2003, 31, 1250-1256).

DIC and SIRS may occur during sepsis, but also as a result ofoperations, tumour disorders, burns or other injuries. In the case ofDIC, there is a massive activation of the coagulation system at thesurface of damaged endothelial cells, the surfaces of foreign bodies orinjured extravascular tissue. As a consequence, there is coagulation insmall vessels of various organs with hypoxia and subsequent organdysfunction. A secondary effect is the consumption of coagulationfactors (for example factor X, prothrombin, fibrinogen) and platelets,which reduces the coagulability of the blood and may result in heavybleeding.

In addition, the inventive compounds can also be used for preventingcoagulation ex vivo, for example for preserving blood and plasmaproducts, for cleaning/pretreating catheters and other medicalauxiliaries and instruments, including extracorporeal circulation, forcoating synthetic surfaces of medical auxiliaries and instruments usedin vivo or ex vivo or for platelet-containing biological samples.

The present invention further provides for the use of the inventivecompounds for coating medical instruments and implants, for examplecatheters, prostheses, stents or artificial heart valves. In this case,the inventive compounds can be firmly attached to the surface or, forlocal action, be released over a certain period of time from a carriercoating into the immediate environment.

The present invention further provides for the use of the inventivecompounds for treatment and/or prophylaxis of disorders, in particularthe disorders mentioned above.

The present invention further provides for the use of the inventivecompounds for producing a medicament for treatment and/or prophylaxis ofdisorders, in particular the disorders mentioned above.

The present invention further provides a method for treatment and/orprophylaxis of disorders, in particular the disorders mentioned above,using a therapeutically effective amount of an inventive compound.

The present invention further provides medicaments comprising aninventive compound and one or more further active ingredients, inparticular for treatment and/or prophylaxis of the disorders mentionedabove. Active ingredients suitable for combinations are, by way ofexample and with preference:

calcium channel blockers, for example amlodipine besilate (for exampleNorvasc®), felodipine, diltiazem, verapamil, nifedipine, nicardipine,nisoldipine and bepridil;iomerizine;statins, for example atorvastatin, fluvastatin, lovastatin,pitavastatin, pravastatin, rosuvastatin and simvastatin;cholesterol resorption inhibitors, for example ezetimibe and AZD4121;cholesteryl ester transfer protein (“CETP”) inhibitors, for exampletorcetrapib;low molecular weight heparins, for example dalteparin sodium, ardeparin,certoparin, enoxaparin, parnaparin, tinzaparin, reviparin andnadroparin;further anticoagulants, for example warfarin, marcumar, fondaparinux;antiarrhythmics, for example dofetilide, ibutilide, metoprolol,metoprolol tartrate, propranolol, atenolol, ajmaline, disopyramide,prajmaline, procainamide, quinidine, sparteine, aprindine, lidocaine,mexiletine, tocamide, encamide, flecamide, lorcamide, moricizine,propafenone, acebutolol, pindolol, amiodarone, bretylium tosylate,bunaftine, sotalol, adenosine, atropine and digoxin;alpha-adrenergic agonists, for example doxazosin mesylate, terazoson andprazosin;beta-adrenergic blockers, for example carvedilol, propranolol, timolol,nadolol, atenolol, metoprolol, bisoprolol, nebivolol, betaxolol,acebutolol and bisoprolol;aldosterone antagonists, for example eplerenone and spironolactone;angiotensin-converting enzyme inhibitors (“ACE inhibitors”), for examplemoexipril, quinapril hydrochloride, ramipril, lisinopril, benazeprilhydrochloride, enalapril, captopril, spirapril, perindopril, fosinopriland trandolapril;angiotensin II receptor blockers (“ARBs”), for exampleolmesartan-medoxomil, candesartan, valsartan, telmisartan, irbesartan,losartan and eprosartan;endothelin antagonists, for example tezosentan, bosentan andsitaxsentan-sodium;inhibitors of neutral endopeptidase, for example candoxatril andecadotril;phosphodiesterase inhibitors, for example milrinone, theophylline,vinpocetine, EHNA (erythro-9-(2-hydroxy-3-nonyl)adenine), sildenafil,vardenafil and tadalafil;fibrinolytics, for example reteplase, alteplase and tenecteplase;GP IIb/IIIa antagonists, for example integrillin, abciximab andtirofiban;direct thrombin inhibitors, for example AZD0837, argatroban, bivalirudinand dabigatran;indirect thrombin inhibitors, for example odiparcil;direct and indirect factor Xa inhibitors, for examplefondaparinux-sodium, apixaban, razaxaban, rivaroxaban (BAY 59-7939),KFA-1982, DX-9065a, AVE3247, otamixaban (XRP0673), AVE6324, SAR377142,idraparinux, SSR126517, DB-772d, DT-831j, YM-150, 813893, LY517717 andDU-1766;direct and indirect factor Xa/IIa inhibitors, for exampleenoxaparin-sodium, AVE5026, SSR128428, SSR128429 and BIBT-986(Tanogitran);lipoprotein-associated phospholipase A2 (“LpPLA2”) modulators;diuretics, for example chlorthalidone, ethacrynic acid, furosemide,amiloride, chlorothiazide, hydrochlorothiazide, methylclothiazide andbenzthiazide;nitrates, for example isosorbide 5-mononitrate;thromboxane antagonists, for example seratrodast, picotamide andramatroban;platelet aggregation inhibitors, for example clopidogrel, tiklopidin,cilostazol, aspirin, abciximab, limaprost, eptifibatide and CT-50547;cyclooxygenase inhibitors, for example meloxicam, rofecoxib andcelecoxib;B-type natriuretic peptides, for example nesiritide and ularitide;NV1FGF modulators, for example XRP0038;HT1B/5-HT2A antagonists, for example SL65.0472;guanylate cyclase activators, for example ataciguat (HMR1766), HMR1069,riociguat and cinaciguat;e-NOS transcription enhancers, for example AVE9488 and AVE3085;antiatherogenic substances, for example AGI-1067:CPU inhibitors, for example AZD9684;renin inhibitors, for example aliskirin and VNP489;inhibitors of adenosine diphosphate-induced platelet aggregation, forexample clopidogrel, ticlopidine, prasugrel, AZD6140, ticagrelor andelinogrel;NHE-1 inhibitors, for example AVE4454 and AVE4890.

Antibiotic therapy: various antibiotics or antifungal medicamentcombinations are suitable, either as calculated therapy (before amicrobial assessment has been made) or as specific therapy; fluidtherapy, for example crystalloid or colloidal fluids; vasopressors, forexample norepinephrine, dopamine or vasopressin; inotropic therapy, forexample dobutamine; corticosteroids, for example hydrocortisone, orfludrocortisone; recombinant human activated protein C, Xigris; bloodproducts, for example erythrocyte concentrates, platelet concentrates,erythropoietin or fresh frozen plasma; assisted ventilation insepsis-induced acute lung injury (ALI) or acute respiratory distresssyndrome (ARDS), for example permissive hypercapnia, low tidal volumes;sedation: for example diazepam, lorazepam, midazolam or propofol.Opioids: for example fentanyl, hydromorphone, morphine, meperidine orremifentanil. NSAIDs: for example ketorolac, ibuprofen or acetaminophen.Neuromuscular blockade: for example pancuronium; glucose control, forexample insulin, glucose; renal replacement therapies, for examplecontinuous veno-venous haemofiltration or intermittent haemodialysis.Low-dose dopamine for renal protection; anticoagulants, for example forthrombosis prophylaxis or for renal replacement therapies, for exampleunfractionated heparins, low molecular weight heparins, heparinoids,hirudin, bivalirudin or argatroban; bicarbonate therapy; stress ulcerprophylaxis, for example H2 receptor inhibitors, antacids.

Medicaments for proliferative disorders: uracil, chlormethine,cyclophosphamide, ifosfamide, melphalan, chlorambucil, pipobroman,triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine,lomustine, streptozocin, dacarbazine, methotrexate, 5-fluorouracil,floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabinephosphate, pentostatin, vinblastine, vincristine, vindesine, bleomycin,dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin,paclitaxel, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase,interferons, etoposide, teniposide, 17.alpha.-ethynylestradiol,diethylstilbestrol, testosterone, prednisone, fluoxymesterone,dromostanolone propionate, testolactone, megestrol acetate, tamoxifen,methylprednisolone, methyltestosterone, prednisolone, triamcinolone,chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estranrustine,medroxyprogesterone acetate, leuprolide, flutamide, toremifene,goserelin, cisplatin, carboplatin, hydroxyurea, amsacrine, procarbazine,mitotane, mitoxantrone, levamisole, navelbene, anastrazole, letrazole,capecitabine, reloxafine, droloxafine, hexamethylmelamine, oxaliplatin(Eloxatin), Iressa (gefmitib, Zd1839), XELODA® (capecitabine), Tarceva®(erlotinib), Azacitidine (5-azacytidine; 5-AzaC), temozolomide(Temodar®), gemcitabine (e.g. GEMZAR® (gemcitabine HCl)), vasostatin ora combination of two or more of the above.

The present invention further provides a method for preventing thecoagulation of blood in vitro, in particular in banked blood orbiological samples containing platelets, which is characterized in thatan anticoagulatory amount of the inventive compound is added.

The inventive compounds can act systemically and/or locally. For thispurpose, they can be administered in a suitable way, for example by theoral, parenteral, pulmonal, nasal, sublingual, lingual, buccal, rectal,dermal, transdermal, conjunctival, otic route, or as implant or stent.

The inventive compounds can be administered in administration formssuitable for these administration routes.

Suitable administration forms for oral administration are those whichfunction according to the prior art and deliver the inventive compoundsrapidly and/or in modified fashion, and which contain the inventivecompounds in crystalline and/or amorphized and/or dissolved form, forexample tablets (uncoated or coated tablets, for example having entericcoatings or coatings which are insoluble or dissolve with a delay andcontrol the release of the inventive compound), tablets whichdisintegrate rapidly in the mouth, or films/wafers, films/lyophilizates,capsules (for example hard or soft gelatin capsules), sugar-coatedtablets, granules, pellets, powders, emulsions, suspensions, aerosols orsolutions.

Parenteral administration can take place with avoidance of an absorptionstep (e.g. intravenous, intraarterial, intracardiac, intraspinal orintralumbar) or with inclusion of an absorption (e.g. intramuscular,subcutaneous, intracutaneous, percutaneous or intraperitoneal).Administration forms suitable for parenteral administration includepreparations for injection and infusion in the form of solutions,suspensions, emulsions, lyophilizates or sterile powders.

Oral administration is preferred.

Suitable for the other administration routes are, for example,pharmaceutical forms for inhalation (inter alia powder inhalers,nebulizers), nasal drops, solutions or sprays; tablets for lingual,sublingual or buccal administration, films/wafers or capsules,suppositories, preparations for the ears or eyes, vaginal capsules,aqueous suspensions (lotions, shaking mixtures), lipophilic suspensions,ointments, creams, transdermal therapeutic systems (e.g. patches), milk,pastes, foams, dusting powders, implants or stents.

The inventive compounds can be converted to the administration formsmentioned. This can be done in a manner known per se by mixing withinert, nontoxic, pharmaceutically suitable excipients. These excipientsinclude carriers (for example microcrystalline cellulose, lactose,mannitol), solvents (e.g. liquid polyethylene glycols), emulsifiers anddispersants or wetting agents (for example sodium dodecylsulphate,polyoxysorbitan oleate), binders (for example polyvinylpyrrolidone),synthetic and natural polymers (for example albumin), stabilizers (e.g.antioxidants for example ascorbic acid), colours (e.g. inorganicpigments for example iron oxides) and masking flavours and/or odours.

The present invention further provides medicaments comprising at leastone inventive compound, preferably together with one or more inertnontoxic pharmaceutically suitable auxiliaries, and their use for thepurposes mentioned above.

In the case of parenteral administration, it has generally been found tobe advantageous to administer amounts of about 5 to 250 mg every 24hours to achieve effective results. In the case of oral administrationthe amount is about 5 to 100 mg every 24 hours.

It may nevertheless be necessary where appropriate to deviate from thestated amounts, in particular as a function of the body weight, route ofadministration, individual response to the active ingredient, nature ofthe preparation and time or interval over which administration takesplace.

The percentages in the tests and examples which follow are, unlessindicated otherwise, percentages by weight; parts are parts by weight.Solvent ratios, dilution ratios and concentration data for theliquid/liquid solutions are each based on volume. “w/v” means“weight/volume”. For example, “10% w/v” means: 100 ml of solution orsuspension comprise 10 g of substance.

A) EXAMPLES Abbreviations

-   approx. approximately-   CDI carbonyldiimidazole-   d day(s), doublet (in NMR)-   TLC thin-layer chromatography-   DCI direct chemical ionization (in MS)-   dd double doublet (in NMR)-   DMAP 4-dimethylaminopyridine-   DMF N,N-dimethylformamide-   DMSO dimethyl sulphoxide-   DPPA diphenyl phosphorazidate-   DSC disuccinimidyl carbonate-   eq. equivalent(s)-   ESI electrospray ionization (in MS)-   h hour(s)-   HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate-   HPLC high-pressure, high-performance liquid chromatography-   LC-MS liquid chromatography-coupled mass spectroscopy-   LDA lithium diisopropylamide-   m multiplet (in NMR)-   min minute(s)-   MS mass spectroscopy-   NMR nuclear magnetic resonance spectroscopy-   PYBOP benzotriazol-1-yloxytris(pyrrolidino)phosphonium    hexafluorophosphate-   q quartet (in NMR)-   RP reversed phase (in HPLC)-   RT room temperature-   R_(t) retention time (in HPLC)-   singlet (in NMR)-   t triplet (in NMR)-   THF tetrahydrofuran

HPLC Methods:

Method 1A:

Instrument: HP 1100 with DAD detection; column: Kromasil 100 RP-18, 60mm×2.1 mm, 3.5 μm; eluent A: 5 ml of perchloric acid (70%)/1 of water,eluent B: acetonitrile; gradient: 0 min 2% B→0.5 min 2% B→4.5 min 90%B→6.5 min 90% B→6.7 min 2% B→7.5 min 2% B; flow rate: 0.75 ml/min;column temperature: 30° C.; UV detection: 210 nm.

LC-MS Methods:

Method 1B:

MS instrument type: Micromass ZQ; HPLC instrument type: HP 1100 Series;UV DAD; column: Phenomenex Gemini 3μ, 30 mm×3.0 mm; eluent A: 1 l ofwater+0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile+0.5 ml of50% formic acid; gradient: 0.0 min 90% A→2.5 min 30% A→3.0 min 5% A→4.5min 5% A; flow rate: 0.0 min 1 ml/min, 2.5 min/3.0 min/4.5 min 2 ml/min;oven: 50° C.; UV detection: 210 nm.

Method 2B:

Instrument: Micromass QuattroPremier with Waters UPLC Acquity; column:Thermo Hypersil GOLD 1.9μ, 50 mm×1 mm; eluent A: 1 l of water+0.5 ml of50% formic acid, eluent B: 1 l of acetonitrile+0.5 ml of 50% formicacid; gradient: 0.0 min 90% A→0.1 min 90% A→1.5 min 10% A→2.2 min 10% A;oven: 50° C.; flow rate: 0.33 ml/min; UV detection: 210 nm.

Method 3B:

MS instrument type: Micromass ZQ; HPLC instrument type: Waters Alliance2795; column: Phenomenex Synergi 2.5μ MAX-RP 100A Mercury, 20 mm×4 mm;eluent A: 1 l of water+0.5 ml of 50% formic acid, eluent B: 1 l ofacetonitrile+0.5 ml of 50% formic acid; gradient: 0.0 min 90% A→0.1 min90% A→3.0 min 5% A→4.0 min 5% A→4.01 min 90% A; flow rate: 2 ml/min;oven: 50° C.; UV detection: 210 nm.

Method 4B:

Instrument: Micromass Quattro Micro MS with HPLC Agilent series 1100;column: Thermo Hypersil GOLD 3μ 20 mm×4 mm; eluent A: 1 l of water+0.5ml of 50% formic acid, eluent B: 1 l of acetonitrile+0.5 ml of 50%formic acid; gradient: 0.0 min 100% A→3.0 min 10% A→4.0 min 10% A→4.01min 100% A→5.00 min 100% A; oven: 50° C.; flow rate: 2 ml/min; UVdetection: 210 nm.

Method 5B:

Instrument: Waters ACQUITY SQD UPLC System; column: Waters Acquity UPLCHSS T3 1.8μ 50 mm×1 mm; eluent A: 1 l of water+0.25 ml of 99% formicacid, eluent B: 1 l of acetonitrile+0.25 ml of 99% formic acid;gradient: 0.0 min 90% A→1.2 min 5% A→2.0 min 5% A; oven: 50° C.; flowrate: 0.40 ml/min; UV detection: 210-400 nm.

Method 6B:

MS instrument type: Waters (Micromass) Quattro Micro; HPLC instrumenttype: Agilent 1100 series; column: Thermo Hypersil GOLD 3μ 20 mm×4 mm;eluent A: 1 l of water+0.5 ml of 50% formic acid, eluent B: 1 l ofacetonitrile+0.5 ml of 50% formic acid; gradient: 0.0 min 100% A→3.0 min10% A→4.0 min 10% A→4.01 min 100% A; (flow rate: 2.5 ml/min)→5.00 min100% A; oven: 50° C.; flow rate: 2 ml/min; UV detection: 210 nm.

Method 7B:

MS instrument type: Waters ZQ; HPLC instrument type: Agilent 1100Series; UV DAD; column: Thermo Hypersil GOLD 3μ 20 mm×4 mm; eluent A: 1l of water+0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile+0.5ml of 50% formic acid; gradient: 0.0 min 100% A→3.0 min 10% A→4.0 min10% A oven: 55° C.; flow rate: 2 ml/min; UV detection: 210 nm.

Preparative Separation of Enantiomers:

Method 1D:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×20 mm, eluent:isohexane/isopropanol 25:75; flow rate: 15 ml/min, temperature: 45° C.;UV detection: 220 nm.

Method 2D:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×20 mm, eluent:methanol/acetonitrile 25:75; flow rate: 15 ml/min, temperature: 30° C.;UV detection: 220 nm.

Method 3D:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×20 mm, eluent:methanol/acetonitrile 50:50; flow rate: 15 ml/min, temperature: 30° C.;UV detection: 220 nm.

Method 4D:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×20 mm, eluent: tert-butyl methylether/methanol 50:50; flow rate: 15 ml/min; temperature: 30° C.; UVdetection: 220 nm.

Method 5D:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×20 mm, eluent:methanol/acetonitrile 25:75; flow rate: 15 ml/min, temperature: 30° C.;UV detection: 220 nm.

Method 6D:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×20 mm, eluent:isohexane/ethanol 25:75; flow rate: 15 ml/min, temperature: 45° C.; UVdetection: 220 nm.

Method 7D:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×20 mm, eluent: ethanol 100%;flow rate: 15 ml/min, temperature: 45° C.; UV detection: 220 nm.

Method 8D:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×20 mm, eluent:isohexane/isopropanol 30:70; flow rate: 15 ml/min, temperature: 45° C.;UV detection: 220 nm.

Method 9D:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×20 mm, eluent:acetonitrile/methanol 70:30; flow rate: 15 ml/min, temperature: 30° C.;UV detection: 220 nm.

Method 10D:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×20 mm, eluent:acetonitrile/methanol 70:30; flow rate: 20 ml/min, temperature: 35° C.;UV detection: 210 nm.

Method 11D:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×20 mm, eluent:isohexane/ethanol 70:30; flow rate: 15 ml/min, temperature: 40° C.; UVdetection: 220 nm.

Analytical Separation of Enantiomers:

Method 1E:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×4 mm; eluent:isopropanol/isohexane: 75:25; flow rate: 1 ml/min; temperature: 45° C.;UV detection: 220 nm.

Method 2E:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×4.6 mm; eluent:isohexane/isopropanol 25:75+0.2% trifluoroacetic acid+1% water; flowrate: 1 ml/min; temperature: 45° C.; UV detection: 235 nm.

Method 3E:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×4.6 mm, eluent:acetonitrile/methanol 75:25; flow rate: 1 ml/min; temperature: 25° C.;UV detection: 220 nm.

Method 4E:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×4.6 mm, eluent:acetonitrile/methanol 50:50; flow rate: 1 ml/min; temperature: 25° C.;UV detection: 220 nm.

Method 5E:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×4.6 mm, eluent: tert-butylmethyl ether/methanol 50:50; flow rate: 1 ml/min; temperature: 25° C.;UV detection: 220 nm.

Method 6E:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×4.6 mm, eluent: ethanol 100%;flow rate: 1 ml/min, temperature: 45° C.; UV detection: 220 nm.

Method 7E:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×4.6 mm, eluent:isohexane/isopropanol 30:70; flow rate: 1 ml/min, temperature: 45° C.;UV detection: 220 nm.

Method 8E:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×4.6 mm, eluent:acetonitrile/methanol 70:30; flow rate: 15 ml/min, temperature: 25° C.;UV detection: 220 nm.

Method 9E:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×4.6 mm, eluent:acetonitrile/methanol 70:30; flow rate: 15 ml/min, temperature: 30° C.;UV detection: 220 nm.

Method 10E:

Phase: Daicel Chiralpak IA, 5 μm 250 mm×4.6 mm, eluent:acetonitrile/methanol 70:30; flow rate: 1 ml/min, temperature: 30° C.;UV detection: 220 nm.

Method 11E:

Phase: Daicel Chiralpak AD-H, 5 μm 250 mm×4.6 mm, eluent:isohexane/ethanol 25:75+0.2% trifluoroacetic acid+1% water; flow rate: 1ml/min, temperature: 45° C.; UV detection: 220 nm.

GC-MS Methods:

Method 1F:

Instrument: Micromass GCT, GC6890; column: Restek RTX-35, 15 m×200μm×0.33 μm; constant flow rate with helium: 0.88 ml/min; oven: 70° C.;inlet: 250° C.; gradient: 70° C., 30° C./min→310° C. (hold for 3 min)

The microwave reactor used was a “single mode” instrument of the Emrys™Optimizer type.

Starting Compounds General Method 1A: N′-Hydroxyimidamide Formation

A solution of the appropriate nitrile (1.0 eq) in ethanol (1.2 ml/mmol)is admixed at RT with hydroxylammonium chloride (1.5 eq.) andtriethylamine (1.2 eq.). The reaction mixture is stirred at roomtemperature overnight. For workup, the ethanol is removed under reducedpressure, saturated aqueous sodium hydrogencarbonate solution is addedand the reaction mixture is extracted with ethyl acetate. The organicphase is dried over sodium sulphate and concentrated. The residue isreacted without further purification.

General Method 2A: N′-Hydroxyimidamide Formation

A solution of the appropriate nitrile (1.0 eq) in a mixture of ethanol(1.9 ml/mmol) and water (0.5 ml/mmol) is admixed at RT withhydroxylammonium chloride (1.08 eq.) and sodium hydroxide (1.12 eq.).The reaction mixture is stirred at room temperature for 16 hours. Forworkup, the reaction mixture is concentrated under reduced pressure,admixed with dichloromethane and filtered. The filtrate is concentratedunder reduced pressure and the residue is reacted without furtherpurification.

General Method 3A: Suzuki Coupling

A mixture of the appropriate bromopyridine in toluene (1.8 ml/mmol) isadmixed under argon and at RT with tetrakis(triphenylphosphine)palladium(0.02 eq.), with a solution of the appropriate arylboronic acid (1.2eq.) in ethanol (0.5 ml/mmol) and with a solution of potassium fluoride(2.0 eq.) in water (0.2 ml/mmol). The reaction mixture is stirred underreflux for several hours until the conversion is substantially complete.After addition of ethyl acetate and phase separation, the organic phaseis washed once with water and once with saturated aqueous sodiumchloride solution, dried (magnesium sulphate), filtered and concentratedunder reduced pressure. The crude product is purified by flashchromatography (silica gel 60, eluent: dichloromethane/methanolmixtures).

General Method 4A: Hydrogenation of the Pyridine

A solution of the pyridine in ethanol (9 ml/mmol) is admixed under argonwith palladium on activated carbon (moistened with approx. 50% water,0.3 g/mmol), and the mixture is hydrogenated at 60° C. in a 50 barhydrogen atmosphere overnight. The catalyst is then filtered off througha filter layer and washed repeatedly with ethanol. The combinedfiltrates are concentrated under reduced pressure.

General Method 5A: Methyl Ester Hydrolysis/Epimerization

At RT, potassium tert-butoxide (10 eq.) is added to a solution of theappropriate methyl ester (1.0 eq.) in methanol (35-40 ml/mmol). Themixture is stirred at 60° C. overnight. If the conversion is incomplete,water (1.0 eq.) is added and the mixture is stirred at 60° C. until theconversion is complete. For workup, the methanol is removed underreduced pressure, the residue is admixed with water and the mixture isacidified (pH 1) with aqueous 1 N hydrochloric acid solution. Themixture is extracted with ethyl acetate and the organic phase is driedwith magnesium sulphate, filtered and concentrated under reducedpressure.

General Method 6A: Oxadiazole Formation

A solution of the appropriate piperidine-3-carboxylic acid indimethylformamide (10-20 ml/mmol) is admixed under argon at RT with HATU(1.2 eq.), N,N-diisopropylethylamine (2.2 eq.) and the appropriateN-hydroxyimidamide (1.1 eq.). The reaction mixture is stirred at RTuntil the formation of the intermediate is complete and then stirredfurther at 120° C. until the desired product has formed from thisintermediate. The reaction mixture is then purified by means ofpreparative HPLC.

Example 1A N′-Hydroxy-3-methoxypropanimidamide

According to General Method 1A, 20.0 g (235.0 mmol) of3-methoxypropionitrile were reacted. Yield: 18.1 g (49% of theory,purity 74%)

HPLC (Method 1A): R_(t)=0.35 min; MS (ESIpos): m/z=119 [M+H]⁺.

Example 2A 3-Ethoxy-N′-hydroxypropanimidamide

According to General Method 2A, 5.0 g (50.4 mmol) of3-ethoxypropionitrile were reacted. Yield: 0.6 g (8% of theory, purity90%)

HPLC (Method 1A): R_(t)=0.60 min; MS (ESIpos): m/z=133 [M+H]⁺.

Example 3A N′-Hydroxycyclopropanecarboximidamide

According to General Method 2A, 7.2 g (107.3 mmol) ofcyclopropanecarbonitrile were reacted. Yield: 4.8 g (44% of theory)

LC-MS (Method 2B): R_(t)=0.16 min; MS (ESIpos): m/z=101 [M+H]⁺.

Example 4A Methyl 5-(4-ethylphenyl)pyridine-3-carboxylate

According to General Method 3A, 32 g (148 mmol) of methyl5-bromonicotinate and 27 g (178 mmol, 1.2 eq.) of 4-ethylphenylboronicacid were reacted. Yield: 24 g (64% of theory)

LC-MS (Method 3B): R_(t)=2.03 min; MS (ESIpos): m/z=242 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=9.13 (d, 1H), 9.05 (d, 1H), 8.45 (t, 1H),7.72 (d, 2H), 7.38 (d, 2H), 3.93 (s, 3H), 2.68 (q, 2H), 1.22 (t, 3H).

Example 5A Methyl 5-(4-ethylphenyl)piperidine-3-carboxylate [racemiccis/trans isomer mixture]

According to General Method 4A, 24 g (94 mmol) of methyl5-(4-ethylphenyl)pyridine-3-carboxylate were hydrogenated. Yield: 20 g(77% of theory)

LC-MS (Method 4B): R_(t)=1.43 min; MS (ESIpos): m/z=248 [M+H]⁺.

Example 6A

Methyl5-(4-ethylphenyl)-1-(thiomorpholin-4-ylcarbonyl)piperidine-3-carboxylate[racemic cis/trans isomer mixture]

5.00 g (12.1 mmol) of 3-methyl 1-(4-nitrophenyl)5-(4-ethylphenyl)piperidine-1,3-dicarboxylate (Example 30A), 3.57 g(36.4 mmol) of thiomorpholine and 5.03 g (36.4 mmol) of potassiumcarbonate were added to 76 ml of DMF and heated in 5 portions at 150° C.for 1.5 h in a single-mode microwave (Emrys Optimizer). For workup, thereaction solutions were combined and filtered, and the residue waspurified by means of preparative HPLC. Yield: 3.07 g (67% of theory)

LC-MS (Method 5B): R_(t)=1.16 and 1.18 min (cis/trans isomers); MS(ESIpos): m/z=377 [M+H]⁺.

Example 7A5-(4-Ethylphenyl)-1-(thiomorpholin-4-ylcarbonyl)piperidine-3-carboxylicacid [racemic cis isomer]

According to General Method 5A, 3.00 g (7.97 mmol) of the compound fromExample 6A and 8.94 g (79.7 mmol) of potassium tert-butoxide werereacted. The reaction led selectively to the cis isomer. Yield: 2.74 g(93% of theory)

LC-MS (Method 5B): R_(t)=1.04 min; MS (ESIpos): m/z=363 [M+H]⁺.

Example 8A{3-(4-Ethylphenyl)-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 300 mg (0.828 mmol) of the compound fromExample 7A and 134 mg (0.910 mmol) ofN′-hydroxy-3-methoxypropanimidamide were reacted Yield: 185 mg (49% oftheory)

LC-MS (Method 5B): R_(t)=1.22 min; MS (ESIpos): m/z=445 [M+H]⁺.

Example 9A[3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-(4-ethylphenyl)piperidin-1-yl](thiomorpholin-4-yl)-methanone[racemic cis isomer]

According to General Method 6A, 300 mg (0.828 mmol) of the compound fromExample 7A and 91 mg (0.91 mmol) of N-hydroxycyclopropanecarboximidamidewere reacted Yield: 141 mg (40% of theory)

LC-MS (Method 5B): R_(t)=1.32 min; MS (ESIpos): m/z=427 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=7.22 (d, 2H), 7.15 (d, 2H), 3.92 (d, 1H),3.52 (d, 1H), 3.44 (br. s., 4H), 3.38-3.31 (m, 1H), 3.03-2.79 (m, 3H),2.63-2.55 (m, 6H), 2.25 (d, 1H), 2.10 (td, 1H), 1.91 (q, 1H), 1.16 (t,3H), 1.09-1.01 (m, 2H), 0.92-0.85 (m, 2H).

Example 10A{3-(4-Ethylphenyl)-5-[3-(2-hydroxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 300 mg (0.828 mmol) of the compound fromExample 7A and 112 mg (1.08 mmol) of N′,3-dihydroxypropanimidamide[Graham A. Showell et al., J. Med. Chem., 1991, 34, 1086-1094] werereacted. Yield: 248 mg (66% of theory)

LC-MS (Method 5B): R_(t)=2.22 min; MS (ESIpos): m/z=431 [M+H]⁺.

Example 11A{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-(4-ethylphenyl)piperidin-1-yl}(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 600 mg (1.655 mmol) of the compound fromExample 7A and 355 mg (approx. 2.152 mmol) of3-Ethoxy-N′-hydroxypropanimidamide were reacted Yield: 389 mg (49% oftheory)

LC-MS (Method 6B): R_(t)=2.61 min; MS (ESIpos): m/z=459 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.16 (d, 2H), 3.95 (d, 1H),3.71 (t, 2H), 3.54 (d, 1H), 3.48-3.34 (m, 7H), 3.08-2.81 (m, 5H),2.63-2.55 (m, 6H), 2.29 (d, 1H), 1.95 (q, 1H), 1.16 (t, 3H), 1.07 (t,3H).

Example 12A Methyl 5-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylate

According to General Method 3A, 28 g (132 mmol) of methyl5-bromonicotinate and 30 g (158 mmol, 1.2 eq.) of4-trifluoromethylphenylboronic acid were reacted. Yield: 32 g (85% oftheory)

LC-MS (Method 4B): R_(t)=2.27 min; MS (ESIpos): m/z=282 [M+H]⁺.

Example 13A Methyl 5-[4-(trifluoromethyl)phenyl]piperidine-3-carboxylate[racemic cis/trans isomer mixture]

32 g (112 mmol) of methyl5-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylate (Example 12A) werehydrogenated according to General Method 4A. Yield: 26 g (82% of theory)

LC-MS (Method 1B): R_(t)=1.35 and 1.41 min (cis/trans isomers); MS(ESIpos): m/z=288 [M+H]±.

¹H NMR (400 MHz, DMSO-d₆): δ=9.22 (d, 1H), 9.14 (d, 1H), 8.57 (t, 1H),8.06 (d, 2H), 7.89 (d, 2H), 3.94 (s, 3H).

Example 14A 3-Methyl 1-(4-nitrophenyl)5-[4-(trifluoromethyl)phenyl]piperidine-1,3-dicarboxylate [racemiccis/trans isomer mixture]

20.0 g (69.6 mmol) of methyl5-[4-(trifluoromethyl)phenyl]piperidine-3-carboxylate (Example 13A) weredissolved in 1.0 l of dichloromethane, and admixed at 0° C. with 14.1 g(139 mmol) of triethylamine. Subsequently, 14.0 g (69.6 mmol) of4-nitrophenyl chlorocarbonate were added dropwise. The reaction mixturewas stirred at 0° C. for 2 h and then at RT for 16 h. For workup, themixture was washed with saturated aqueous sodium hydrogencarbonatesolution. The organic phase was dried over magnesium sulphate, filteredand concentrated under reduced pressure. This gave 31.3 g of crudeproduct, which was reacted without any further purification steps.

LC-MS (Method 3B): R_(t)=2.44 min and 2.48 min (cis/trans isomers); MS(ESIpos): m/z=453 [M+H]±.

Example 15A

Methyl1-(thiomorpholin-4-ylcarbonyl)-5-[4-(trifluoromethyl)phenyl]piperidine-3-carboxylate[racemic cis/trans isomer mixture]

10.0 g (22.1 mmol) of 3-methyl1-(4-nitrophenyl)-5-[4-(trifluoromethyl)phenyl]piperidine-1,3-dicarboxylate,6.84 g (66.3 mmol) of thiomorpholine and 9.17 g (66.3 mmol) of potassiumcarbonate were added to 150 ml of DMF and heated in 10 portions at 150°C. for 1 h in a single-mode microwave (Emrys Optimizer). For workup, thereaction solutions were combined and filtered, and the residue waspurified by means of preparative HPLC. Yield: 5.16 g (55% of theory)

LC-MS (Method 5B): R_(t)=1.13 and 1.16 min (cis/trans isomers); MS(ESIpos): m/z=417 [M+H]±.

Example 16A1-(Thiomorpholin-4-ylcarbonyl)-5-[4-(trifluoromethyl)phenyl]piperidine-3-carboxylicacid [racemic cis isomer]

According to General Method 5A, 5.16 g (12.4 mmol) of the compound fromExample 15A and 13.9 g (124 mmol) of potassium tert-butoxide werereacted. The reaction led selectively to the cis isomer. Yield: 4.90 g(98% of theory)

LC-MS (Method 5B): R_(t)=1.04 min; MS (ESIpos): m/z=403 [M+H]⁺.

Example 17A{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 600 mg (1.491 mmol) of the compound fromExample 16A and 164 mg (1.640 mmol) ofN-hydroxycyclopropanecarboximidamide were reacted Yield: 352 mg (47% oftheory)

LC-MS (Method 5B): R_(t)=1.28 min; MS (ESIpos): m/z=467 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.56 (d, 2H), 3.92 (d, 1H),3.57 (d, 1H), 3.45 (br. s., 4H), 3.40-3.34 (m, 1H), 3.08-2.95 (m, 3H),2.59 (br. s., 4H), 2.30 (d, 1H), 2.16-2.07 (m, 1H), 2.04-1.91 (m, 1H),1.10-1.01 (m, 2H), 0.92-0.85 (m, 2H).

Example 18A{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 600 mg (1.491 mmol) of the compound fromExample 16A and 242 mg (1.640 mmol) ofN′-hydroxy-3-methoxypropanimidamide were reacted Yield: 350 mg (46% oftheory)

LC-MS (Method 5B): R_(t)=1.18 min; MS (ESIpos): m/z=485 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): 6=7.70 (d, 2H), 7.57 (d, 2H), 3.95 (d, 1H),3.68 (t, 2H), 3.58 (d, 1H), 3.51-3.36 (m, 5H), 3.23 (s, 3H), 3.13-2.96(m, 3H), 2.94 (t, 2H), 2.60 (br. s., 4H), 2.33 (br. d., 1H), 2.10-1.95(m, 1H).

Example 19A{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 600 mg (1.491 mmol) of the compound fromExample 16A and 320 mg (approx. 1.983 mmol) of3-ethoxy-N′-hydroxypropanimidamide were reacted Yield: 343 mg (46% oftheory)

LC-MS (Method 6B): R_(t)=2.57 min; MS (ESIpos): m/z=499 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.57 (d, 2H), 3.96 (d, 1H),3.71 (t, 2H), 3.58 (d, 1H), 3.49-3.37 (m, 7H), 3.11-2.97 (m, 3H), 2.93(t, 2H), 2.60 (br. s., 4H), 2.34 (br. d., 1H), 2.02 (q, 1H), 1.07 (t,3H).

Example 20A{3-[3-(2-Hydroxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl}piperidin-1-yl]-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 600 mg (1.491 mmol) of the compound fromExample 16A and 201 mg (1.938 mmol) of N′,3-dihydroxypropanimidamidewere reacted Yield: 494 mg (68% of theory)

LC-MS (Method 5B): R_(t)=1.04 min; MS (ESIpos): m/z=471 [M+H]⁺.

Example 21A Methyl 5-[4-(trifluoromethoxy)phenyl]pyridine-3-carboxylate

According to General Method 3A, 23 g (105 mmol) of methyl5-bromonicotinate and 26 g (126 mmol, 1.2 eq.) of4-trifluoromethoxyphenylboronic acid were reacted. Yield: 14 g (41% oftheory)

LC-MS (Method 1B): R_(t)=2.44 min; MS (ESIpos): m/z=298 [M+H]⁺.

Alternative Synthesis:

A solution of 26 g (121 mmol) of methyl 5-bromonicotinate in toluene(220 ml) was admixed under argon at RT with 2.8 g (2.4 mmol) oftetrakis(triphenylphosphine)palladium, and then a solution of 30 g (146mmol) of 4-trifluoromethoxyphenylboronic acid in ethanol (58 ml) wasadded. After adding 14 g (243 mmol) of potassium fluoride in water (58ml), the mixture was stirred under reflux overnight, a further 0.70 g(0.61 mmol) of tetrakis(triphenylphosphine)palladium was added, and themixture was stirred under reflux for a further 24 h. After addinganother 1.4 g (1.2 mmol) of tetrakis(triphenylphosphine)palladium, themixture was stirred under reflux for 20 h, and the reaction solution wasadmixed with ethyl acetate and washed with water and saturated aqueoussodium chloride solution. The organic phase was dried over magnesiumsulphate, filtered and concentrated under reduced pressure. The residuewas purified by means of column chromatography (silica gel,cyclohexane/dichloromethane 1:1→dichloromethane). Yield: 31 g (86% oftheory)

LC-MS (Method 4B): R_(t)=2.32 min; MS (ESIpos): m/z=298 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=9.17 (d, 1H), 9.10 (d, 1H), 8.51 (t, 1H),7.95 (d, 2H), 7.52 (d, 2H), 3.94 (s, 3H).

Example 22A Methyl5-[4-(trifluoromethoxy)phenyl]piperidine-3-carboxylate [racemiccis/trans isomer mixture]

14 g (45 mmol) of methyl5-[4-(trifluoromethoxy)phenyl]pyridine-3-carboxylate in ethanol (500 ml)were admixed with 17 g of moistened palladium/carbon catalyst (10%palladium, 50% water), and then hydrogenated at 60° C. and a 50 barhydrogen atmosphere overnight. The reaction solution was filtered, thefilter residue was washed with ethanol and the filtrate was concentratedunder reduced pressure. The residue was purified by means of columnchromatography (silica gel, dichloromethane/methanol 600:1→10:1). Yield:8 g (59% of theory)

LC-MS (Method 1B): R_(t)=1.29 min and 1.33 min (cis/trans isomers); MS(ESIpos): m/z=304 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.43-7.35 (m, 4H), 7.31-7.25 (m, 4H), 3.60(s, 3H), 3.40-3.21 (m, 5H), 3.16 (d, 1H), 3.01-2.89 (m, 3H), 2.88-2.78(m, 2H), 2.78-2.65 (m, 4H), 2.17 (d, 1H), 2.09 (d, 1H), 1.82 (td, 1H),1.68 (q, 1H), approx. 1:1.3 mixture of the cis/trans isomers, twoprotons hidden.

Example 23A 3-Methyl1-(4-nitrophenyl)-5-[4-(trifluoromethoxy)phenyl]piperidine-1,3-dicarboxylate[racemic cis/trans isomer mixture]

At 0° C., 5.32 g (26.4 mmol) of 4-nitrophenyl chloroformate were addedslowly to 8.0 g (26.4 mmol) of methyl5-(4-(trifluoromethoxy)phenyl)piperidine-3-carboxylate (Example 22A) and5.34 g (26.3 mmol) of triethylamine in 666 ml of dichloromethane. Themixture was stirred at RT for 2 h. For workup, the reaction mixture waswashed first with saturated aqueous sodium hydrogencarbonate solution,then with water. The organic phase was dried over sodium sulphate andconcentrated under reduced pressure. The residue was purified by meansof flash chromatography on silica gel (eluent: cyclohexane/ethyl acetate1:2->1:1). Yield: 7.32 g (54% of theory)

LC-MS (Method 3B): R_(t)=2.47 min; MS (ESIpos): m/z=469 [M+H]⁺.

Example 24A Methyl1-(thiomorpholin-4-ylcarbonyl)-5-[4-(trifluoromethoxy)phenyl]piperidine-3-carboxylate[racemic cis/trans isomer mixture]

12.0 g (25.1 mmol) of 3-methyl1-(4-nitrophenyl)-5-[4-(trifluoromethoxy)phenyl]piperidine-1,3-dicarboxylate,7.77 g (75.3 mmol) of thiomorpholine and 10.4 g (75.3 mmol) of potassiumcarbonate were added to 180 ml of DMF and heated in 12 portions at 150°C. for 2 h in a single-mode microwave (Emrys Optimizer). For workup, thereaction solutions were combined and filtered, and the residue waspurified by means of preparative HPLC. Yield: 7.88 g (73% of theory)

LC-MS (Method 5B): R_(t)=1.16 and 1.18 min (cis/trans isomers); MS(ESIpos): m/z=433 [M+H]±.

¹H NMR (400 MHz, DMSO-d₆): δ=7.46-7.39 (m, 4H), 7.32 (d, 4H), 3.84 (dd,2H), 3.64 (s, 3H), 3.63 (s, 3H), 3.55-3.34 (m, 10H), 3.09 (dd, 1H),3.06-2.96 (m, 1H), 2.92-2.81 (m, 6H), 2.76-2.67 (m, 1H), 2.65-2.56 (m,7H), 2.25-2.10 (m, 2H), 1.95-1.84 (m, 1H), 1.76 (q, 1H), approx. 1:1mixture of the cis/trans isomers.

Example 25A1-(Thiomorpholin-4-ylcarbonyl)-5-[4-(trifluoromethoxy)phenyl]piperidine-3-carboxylicacid [racemic cis/trans isomer mixture]

20.4 g (182 mmol) of potassium tert-butoxide were added at RT to asolution of 7.85 g (18.2 mmol) of the compound from Example 24A inmethanol (650 ml). The mixture was stirred at 60° C. overnight. Forworkup, the methanol was removed under reduced pressure, the residue wasadmixed with water and the mixture was acidified (pH 1) with aqueous 1 Nhydrochloric acid solution. The mixture was extracted with ethylacetate, and the organic phase was dried with magnesium sulphate,filtered and concentrated under reduced pressure. The reaction led to an85:15 cis/trans isomer mixture. Yield: 7.70 g (99% of theory)

LC-MS (Method 5B): R_(t)=1.03 (trans isomer) and 1.04 min (cis isomer);MS (ESIpos): m/z=419 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=12.44 (br. s., 1H), 7.47-7.39 (m, 2H), 7.31(d, 2H), 3.79 (d, 1H), 3.56-3.48 (m, 1H), 3.46-3.37 (m, 4H), 2.91-2.73(m, 3H), 2.63-2.55 (m, 5H), 2.14 (d, 1H), 1.81-1.66 (m, 1H).

Example 26A{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 600 mg (1.43 mmol) of the compound fromExample 25A and 232 mg (1.58 mmol) ofN′-hydroxy-3-methoxypropanimidamide were reacted Yield: 398 mg (53% oftheory)

LC-MS (Method 5B): R_(t)=1.21 min; MS (ESIpos): m/z=501 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=7.47 (d, 2H), 7.33 (d, 2H), 3.95 (d, 1H),3.68 (t, 2H), 3.56 (d, 1H), 3.50-3.35 (m, 5H), 3.23 (s, 3H), 3.08-2.86(m, 5H), 2.60 (br. s., 4H), 2.32 (d, 1H), 1.97 (q, 3H).

Example 27A{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 300 mg (0.717 mmol) of the compound fromExample 25A and 79 mg (0.789 mmol) ofN-hydroxycyclopropanecarboximidamide were reacted Yield: 135 mg (39% oftheory)

LC-MS (Method 2B): R_(t)=1.44 min; MS (ESIpos): m/z=483 [M+H]⁺.

Alternative Synthesis:

600 mg (1.43 mmol) of the compound from Example 25A in dimethylformamide(29.0 ml) were admixed at RT with 654 mg (1.72 mmol) of HATU and 0.55 ml(498 mg, 3.16 mmol) of N,N-diisopropylethylamine, and the mixture wasstirred for 30 min Subsequently, 158 mg (1.58 mmol) ofN′-hydroxycyclopropanecarboximidamide were added and the mixture wasstirred at RT overnight. The reaction solution was heated to 120° C. andstirred at this temperature for 1 h. The reaction solution wassubsequently purified directly by means of preparative HPLC. Yield: 315mg (45% of theory)

LC-MS (Method 5B): R_(t)=1.30 min; MS (ESIpos): m/z=483 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): 6=7.46 (d, 2H), 7.33 (d, 2H), 3.91 (d, 1H),3.55 (d, 1H), 3.45 (br. s., 4H), 3.39-3.32 (m, 1H), 3.05-2.91 (m, 3H),2.59 (br. s., 4H), 2.28 (d, 1H), 2.17-2.08 (m, 1H), 1.93 (q, 1H),1.10-1.02 (m, 2H), 0.92-0.84 (m, 2H).

Example 28A{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 600 mg (1.434 mmol) of the compound fromExample 25A and 307 mg (approx. 1.864 mmol) of3-ethoxy-N′-hydroxypropanimidamide were reacted Yield: 403 mg (55% oftheory)

LC-MS (Method 6B): R_(t)=2.61 min; MS (ESIpos): m/z=515 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): 6=7.47 (d, 2H), 7.33 (d, 2H), 3.95 (d, 1H),3.71 (t, 2H), 3.56 (d, 1H), 3.50-3.35 (t, 7H), 3.10-2.88 (m, 5H), 2.60(br. s., 4H), 2.32 (d, 1H), 2.02-1.92 (m, 1H), 1.07 (t, 3H).

Example 29A{3-[3-(2-Hydroxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 1.00 g (2.390 mmol) of the compound fromExample 25A and 323 mg (3.107 mmol) of N′,3-dihydroxypropanimidamidewere reacted Yield: 848 mg (69% of theory)

LC-MS (Method 6B): R_(t)=2.26 min; MS (ESIpos): m/z=487 [M+H]⁺.

Example 30A 3-Methyl 1-(4-nitrophenyl)5-(4-ethylphenyl)piperidine-1,3-dicarboxylate [racemic cis/trans isomermixture]

3.0 g (12.1 mmol) of the compound from Example 5A were initially chargedin 30 ml of dichloromethane, cooled to 0° C. and admixed with 3.4 ml(2.4 g, 12.1 mmol) of triethylamine and 2.4 g (12.1 mmol) of4-nitrophenyl chloroformate. The reaction mixture was allowed to warm upslowly to RT and stirred at RT for 16 h. The mixture was washed severaltimes with water, dried over sodium sulphate, filtered and concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (eluentdichloromethane→dichloromethane/methanol 100:2). Yield: 4.7 g (83% oftheory, purity 89%)

HPLC (Method 1A): R_(t)=4.94 min and 5.00 min (cis/trans isomer); MS(ESIpos): m/z=413 [M+H]⁺.

Example 31A 4-Nitrophenyl thiomorpholine-4-carboxylate

7.7 g (74.4 mmol) of thiomorpholine were initially charged in 100 ml ofdichloromethane and, while cooling with an ice bath, admixed with 20.7ml (15.1 g, 148.8 mmol) of triethylamine. 10.0 g (49.6 mmol) of4-nitrophenyl chloroformate were added in portions. The reaction mixturewas stirred at RT for one hour, and admixed with water and ethylacetate. The organic phase was removed, washed with 1 N hydrochloricacid and saturated aqueous sodium chloride solution, dried over sodiumsulphate, filtered and concentrated under reduced pressure. Yield: 13.2g (99% of theory)

LC-MS (Method 5B): R_(t)=0.98 min; MS (ESIpos): m/z=269 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=8.28 (d, 2H), 7.46 (d, 2H), 3.86 (br. s.,2H), 3.72 (br. s., 2H), 2.71 (br. d., 4H).

Example 32A 4-Nitrophenyl thiomorpholine-4-carboxylate 1-oxide

13.1 g (49.0 mmol) of 4-nitrophenyl thiomorpholine-4-carboxylate wereinitially charged in 135 ml of dichloromethane and admixed at 0° C. with7.6 g (44.1 mmol) of m-chloroperbenzoic acid in portions. The mixturewas stirred at RT for two hours, water was added and the organic phasewas removed. The organic phase was washed rapidly with saturated aqueoussodium hydrogencarbonate solution, filtered and concentrated underreduced pressure. The crude product was purified by means of preparativeHPLC. Yield: 7.8 g (56% of theory)

LC-MS (Method 5B): R_(t)=0.69 min; MS (ESIpos): m/z=285 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=8.30 (d, 2H), 7.49 (d, 2H), 4.20-3.70 (m,4H), 3.03 (dt, 2H), 2.85 (d, 2H).

Example 33A [5-(Methoxycarbonyl)pyridin-3-yl]boronic acid hydrochloride

17.6 g (81.4 mmol) of methyl 5-bromonicotinate were initially charged in375 ml of DMF under argon and admixed with 26.9 g (105.8 mmol) of4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane, 3.0 g (3.6mmol) of tris(dibenzylideneacetone)dipalladium(0), 1.8 g (6.5 mmol) oftricyclohexylphosphine and 32.0 mmol (325.9 mmol) of potassium acetate.The reaction mixture was stirred at 100° C. for 20 h. Subsequently, thesolvent was removed under reduced pressure, the residue was admixed with40 ml of water and 140 ml of tert-butyl methyl ether, and the organicphase was removed. The aqueous phase was extracted three times with 80ml each time of tert-butyl methyl ether. The combined organic extractswere washed with saturated aqueous sodium chloride solution, dried overmagnesium sulphate, filtered and concentrated. The residue was taken upin 360 ml of methanol and admixed with 36 ml of concentratedhydrochloric acid. The reaction mixture was heated to reflux for 22 hand then stirred at RT for 12 h. About half of the solvent was removedunder reduced pressure, and the solution was filtered and concentratedfurther under reduced pressure. The oily residue was recrystallizedtwice from acetone, and the residue was taken up in 10 ml of acetone andadmixed with 100 ml of tert-butyl methyl ether. After 16 h, theprecipitate formed was removed from the solution. This precipitate wasstirred in 50 ml of acetone and left to stand at RT for 5 weeks, and thesolution was removed again. The solutions were combined, concentratedand dissolved in 50 ml of tert-butyl methyl ether. The mixture was leftto stand at RT for 5 weeks and then the precipitate was removed. Theprecipitate was washed three times with tert-butyl methyl ether anddried in a drying cabinet under reduced pressure.

LC-MS (Method 4B): R_(t)=0.91 min; MS (ESIpos): m/z=182 [M+H]⁺.

Example 34A Methyl 5-[4-(difluoromethoxy)phenyl]nicotinate

10.0 g (44.8 mmol) of 4-(difluoromethoxy)bromobenzene were reactedaccording to General Method 3A with 14.6 g (67.3 mmol) of[5-(methoxycarbonyl)pyridin-3-yl]boronic acid hydrochloride. The releaseof the hydrochloride was achieved by additional addition of 6.80 g (49.3mmol) of potassium carbonate. Yield: 8.6 g (67% of theory)

LC-MS (Method 2B): R_(t)=1.15 min; MS (ESIpos): m/z=280 [M+H]⁺.

Example 35A

Methyl 5-[4-(difluoromethoxy)phenyl]piperidine-3-carboxylate [racemiccis/trans isomer mixture]

A solution of 8.6 g (30.9 mmol) of methyl5-[4-(difluoromethoxy)phenyl]nicotinate in concentrated acetic acid (112ml) was admixed with 841 mg of palladium/carbon (10% palladium) and 1.12g of platinum(IV) oxide. This was followed by hydrogenation under ahydrogen atmosphere at standard pressure for 24 h. The reaction solutionwas concentrated under reduced pressure. The residue was taken up inwater, acidified (pH 1) with 1 N hydrochloric acid, extracted withdiethyl ether, then basified (pH>10) with saturated aqueous sodiumhydrogencarbonate solution and extracted repeatedly with ethyl acetate.The combined filtrates were dried over sodium sulphate, filtered andconcentrated under reduced pressure. Yield: 6.6 g (74% of theory)

LC-MS (Method 5B): R_(t)=0.65 min and 0.66 min (cis/trans isomers); MS(ESIpos): m/z=286 [M+H]⁺.

Example 36A

Methyl5-[4-(difluoromethoxy)phenyl]-1-[(1,1-dioxidothiomorpholin-4-yl)carbonyl]piperidine-3-carboxylate[cis/trans isomer mixture]

2.2 g (7.7 mmol) of methyl5-[4-(difluoromethoxy)phenyl]piperidine-3-carboxylate were dissolved in14 ml of N-methylpyrrolidone, and admixed with 4.0 ml (3.0 g, 23.0 mmol)of N,N-diisopropylethylamine and 3.5 g (11.5 mmol) of 4-nitrophenylthiomorpholine-4-carboxylate 1,1-dioxide. The reaction mixture wasconverted in a microwave at 180° C. for seven minutes. Subsequently,water and ethyl acetate were added, and the aqueous phase was removedand extracted repeatedly with ethyl acetate. The combined organicextracts were washed with water and saturated aqueous sodium chloridesolution, dried over sodium sulphate, filtered and concentrated underreduced pressure. The residue was taken up in diethyl ether andfiltered, and the filtrate was purified by means of preparative HPLC.Yield: 2.0 g (51% of theory)

LC-MS (Method 5B): R_(t)=0.92 min and 0.94 min (cis/trans isomers); MS(ESIpos): m/z=447 [M+H]⁺.

Example 37A5-[4-(Difluoromethoxy)phenyl]-1-[(1,1-dioxidothiomorpholin-4-yl)carbonyl]piperidine-3-carboxylicacid [racemic cis isomer mixture]

According to General Method 4A, 2.7 g (6.1 mmol) of methyl5-[4-(difluoromethoxy)phenyl]-1-[(1,1-dioxidothiomorpholin-4-yl)carbonyl]piperidine-3-carboxylatewere reacted with 6.9 g (61.3 mmol) of potassium tert-butoxide. Yield:2.1 g (77% of theory)

LC-MS (Method 5B): R_(t)=0.82 min; MS (ESIpos): m/z=433 [M+H]⁺.

Example 38A

Methyl5-[4-(difluoromethoxy)phenyl]-1-[(1-oxidothiomorpholin-4-yl)carbonyl]piperidine-3-carboxylate[cis/trans isomer mixture]

2.2 g (7.7 mmol) of methyl5-[4-(difluoromethoxy)phenyl]piperidine-3-carboxylate were dissolved in14 ml of N-methylpyrrolidone, and admixed with 4.0 ml (3.0 g, 23.0 mmol)of N,N-diisopropylethylamine and 3.3 g (11.5 mmol) of 4-nitrophenylthiomorpholine-4-carboxylate 1-oxide. The reaction mixture was convertedin a microwave at 180° C. for seven minutes. Subsequently, water andethyl acetate were added, and the aqueous phase was removed andextracted repeatedly with ethyl acetate. The combined organic extractswere washed with water and saturated aqueous sodium chloride solution,dried over sodium sulphate, filtered and concentrated under reducedpressure. The residue was purified by means of preparative HPLC. Yield:2.2 g (59% of theory)

LC-MS (Method 5B): R_(t)=0.90 min and 0.92 min (cis/trans isomers); MS(ESIpos): m/z=431 [M+H]⁺.

Example 39A5-[4-(Difluoromethoxy)phenyl]-1-[(1-oxidothiomorpholin-4-yl)carbonyl]piperidine-3-carboxylicacid [racemic cis isomer mixture]

According to General Method 4A, 2.7 g (6.3 mmol) of methyl5-[4-(difluoromethoxy)phenyl]-1-[(1-oxidothiomorpholin-4-yl)carbonyl]piperidine-3-carboxylatewere reacted with 7.1 g (63.3 mmol) of potassium tert-butoxide. Thereaction mixture was concentrated under reduced pressure, and theresidue was suspended in water and acidified with concentratedhydrochloric acid. The precipitate was filtered off, washed with waterand dried under reduced pressure. Yield: 1.1 g (34% of theory)

LC-MS (Method 5B): R_(t)=0.75 min; MS (ESIpos): m/z=417 [M+H]⁺.

Example 40A 1-Bromo-4-(2,2,2-trifluoroethyl)benzene

A solution of 25.0 g (100 mmol) of 4-bromobenzyl bromide in1-methyl-2-pyrrolidone (121 ml) was admixed at RT with 4.95 g (26.0mmol) of copper(I) iodide and 37.5 g (195 mmol) of methyl2,2-difluoro-2-(fluorosulphonyl)acetate. The mixture was heated to 80°C. and then stirred overnight. The reaction solution was added to waterand extracted with diethyl ether, and the organic phase was dried oversodium sulphate. After filtering and concentrating the organic phaseunder reduced pressure, the residue was purified by means of columnchromatography (silica gel, cyclohexane/ethyl acetate 20:1). Yield: 16.1g (67% of theory)

GC-MS (Method 1F): R_(t)=2.66 min; MS (ESIpos): m/z=240 [M+H]⁺.

Example 41A Methyl 5-[4-(2,2,2-trifluoroethyl)phenyl]nicotinate

A solution of 8.00 g (33.5 mmol) of the compound from Example 40A intoluene (304 ml) was admixed under argon at RT with 10.9 g (50.2 mmol)of the compound from Example 33A in ethanol (100 ml) and 5.10 g (36.8mmol) of potassium carbonate. After stirring for 10 min, 3.87 g (3.35mmol) of tetrakis(triphenylphosphine)palladium and then 5.83 g (100mmol) of potassium fluoride in water (64 ml) were added. The mixture wasstirred under reflux for 8 h, and the reaction solution was cooled anddiluted with ethyl acetate. The reaction solution was washed in water,and the organic phase was dried over magnesium sulphate, filtered andconcentrated under reduced pressure. The residue was purified by meansof column chromatography (silica gel, dichloromethane/methanol100:1→80:1). Yield: 9.20 g (69% of theory, purity 75%)

LC-MS (Method 5B): R_(t)=1.06 min; MS (ESIpos): m/z=296 [M+H]⁺.

Example 42A Methyl5-[4-(2,2,2-trifluoroethyl)phenyl]piperidine-3-carboxylate [racemiccis/trans isomer mixture]

A solution of 9.20 g (23.4 mmol) of the compound from Example 41A inconcentrated acetic acid (192 ml) was admixed with 1.94 g ofpalladium/carbon (10% palladium) and 2.23 g of platinum(IV) oxide. Thiswas followed by hydrogenation under a hydrogen atmosphere at standardpressure for 6 h, then addition of another 1.00 g of palladium/carbon(10% palladium) and 2.00 g of platinum(IV) oxide, and hydrogenationunder a hydrogen atmosphere at standard pressure overnight.Subsequently, a further 1.00 g of palladium/carbon (10% palladium) and3.00 g of platinum(IV) oxide were added, and hydrogenation was effectedunder a hydrogen atmosphere at standard pressure for a further 24 h. Thereaction solution was filtered through Celite, the filter residue waswashed with methanol/water and the combined filtrates were concentratedunder reduced pressure. The residue was taken up in dichloromethane andthen washed with a 1 N aqueous sodium carbonate solution. The organicphase was dried over sodium sulphate, filtered and concentrated underreduced pressure. Yield: 6.64 g (85% of theory, purity 90%)

LC-MS (Method 2B): R_(t)=0.83 and 0.84 min (cis/trans isomers); MS(ESIpos): m/z=302 [M+H]⁺.

Example 43A 3-Methyl 1-(4-nitrophenyl)5-[4-(2,2,2-trifluoroethyl)phenyl]piperidine-1,3-dicarboxylate [racemiccis/trans isomer mixture]

A solution of 6.62 g (19.8 mmol, purity 90%) of the compound fromExample 42A in dichloromethane (211 ml) was admixed with 9.65 ml (7.00g, 69.2 mmol) of triethylamine and then admixed at 0° C. with 3.99 g(19.8 mmol) of 4-nitrophenyl chloroformate. The mixture was warmed to RTand stirred for 1 h. The reaction solution was washed with saturatedaqueous sodium hydrogencarbonate solution and water, and the organicphase was dried over magnesium sulphate, filtered and concentrated underreduced pressure. Yield: 10.3 g (91% of theory, purity 81%)

LC-MS (Method 2B): R_(t)=1.40 and 1.42 min (cis/trans isomers); MS(ESIpos): m/z=467 [M+H]⁺.

Example 44A Methyl1-(thiomorpholin-4-ylcarbonyl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidine-3-carboxylate[racemic cis/trans isomer mixture]

A solution of 10.3 g (17.9 mmol, purity 81%) of the compound fromExample 43A in 1-methyl-2-pyrrolidone (65 ml) was admixed with 12.6 ml(13.7 g, 132 mmol) of thiomorpholine and 11.5 ml (8.56 g, 66.2 mmol) ofN,N-diisopropylethylamine and then heated in 5 portions in a single-modemicrowave (Emrys Optimizer) at 150° C. for 1 h. For workup, the reactionsolutions were combined and purified directly by means of preparativeHPLC. Yield: 5.63 g (71% of theory)

LC-MS (Method 5B): R_(t)=1.13 and 1.16 min (cis/trans isomers); MS(ESIpos): m/z=431 [M+H]±.

Example 45A1-(Thiomorpholin-4-ylcarbonyl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidine-3-carboxylicacid [racemic cis isomer]

7.74 g (69.0 mmol) of potassium tert-butoxide were added at RT to asolution of 2.97 g (6.90 mmol) of the compound from Example 44A inmethanol (83 ml). The mixture was stirred at 60° C. overnight. Forworkup, the methanol was removed under reduced pressure, the residue wasadmixed with water and the mixture was acidified (pH 1) with aqueous 1 Nhydrochloric acid solution. The mixture was extracted with ethylacetate, and the organic phase was dried with magnesium sulphate,filtered and concentrated under reduced pressure. Yield: 2.61 g (76% oftheory, purity 84%)

LC-MS (Method 5B): R_(t)=1.02 min; MS (ESIpos): m/z=417 [M+H]⁺.

Example 46A{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 300 mg (0.720 mmol) of the compound fromExample 45A and 79.3 mg (0.792 mmol) ofN′-hydroxycyclopropanecarboximidamide were reacted Yield: 160 mg (45% oftheory)

LC-MS (Method 5B): R_(t)=1.23 min; MS (ESIpos): m/z=481 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=7.32 (s, 4H), 3.92 (d, 1H), 3.68-3.52 (m,3H), 3.44 (br. s., 4H), 3.39-3.33 (m, 1H), 3.03-2.85 (m, 3H), 2.59 (br.s., 5H), 2.28 (d, 1H), 2.16-2.06 (m, 1H), 1.92 (q, 1H), 1.10-1.01 (m,2H), 0.92-0.85 (m, 2H).

Example 47A{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 300 mg (0.720 mmol) of the compound fromExample 45A and 93.6 mg (0.792 mmol) ofN′-hydroxy-3-methoxypropanimidamide were reacted Yield: 231 mg (63% oftheory, approx. 15% trans isomer)

LC-MS (Method 5B): R_(t)=1.14 min; MS (ESIpos): m/z=499 [M+H]⁺.

Example 48A 1-Bromo-4-(1,1-difluoroethyl)benzene

A solution of 10.0 g (50.2 mmol) of 4-bromoacetophenone intetrahydrofuran (20 ml) was admixed with 50.0 ml (151 mmol, 50% intetrahydrofuran) of bis(2-methoxyethyl)aminosulphur trifluoride(Deoxofluor) and 3 drops of methanol, and then stirred under reflux for4 days. The reaction mixture was cautiously added dropwise to a mixtureof saturated aqueous sodium hydrogencarbonate solution and ice (1:1) andthen extracted with diethyl ether. The organic phase was dried oversodium sulphate, filtered and concentrated under reduced pressure. Theresidue was purified by means of column chromatography (silica gel,petroleum ether/dichloromethane 3:1). Yield: 8.46 g (76% of theory)

¹H NMR (400 MHz, DMSO-d₆): 6=7.70 (d, 2H), 7.52 (d, 2H), 1.96 (t, 3H).

Example 49A Methyl 5-[4-(1,1-difluoroethyl)phenyl]nicotinate

A solution of 2.98 g (13.3 mmol) of the compound from Example 48A intoluene (25.0 ml) was admixed under argon at RT with 3.62 g (16.7 mmol)of the compound from Example 33A in ethanol (8.4 ml) and 2.03 g (14.7mmol) of potassium carbonate. After stirring for 10 min, 1.54 g (1.34mmol) of tetrakis(triphenylphosphine)palladium and then 2.33 g (40.0mmol) of potassium fluoride in water (5.8 ml) were added. The mixturewas stirred under reflux for 8 h, and the reaction solution was cooledand diluted with ethyl acetate. The reaction solution was washed inwater, and the organic phase was dried over magnesium sulphate, filteredand concentrated under reduced pressure. The residue was purified bymeans of column chromatography (silica gel, dichloromethane/methanol100:1→80:1). Yield: 2.62 g (69% of theory, 4:1 mixture of methyl andethyl ester)

LC-MS (Method 2B): R_(t)=1.20 min (methyl ester) and 1.28 min (ethylester); MS (ESIpos): m/z=278 [M+H]+(methyl ester) and 292 [M+H]+(ethylester).

Example 50A Methyl5-[4-(1,1-difluoroethyl)phenyl]piperidine-3-carboxylate [racemiccis/trans isomer mixture]

A solution of 2.30 g (8.30 mmol) of the compound from Example 49A inmethanol (52 ml) and concentrated hydrogen chloride solution (6.5 ml)was admixed with 1.05 g of palladium/carbon (10% palladium) and 1.92 gof platinum(IV) oxide and then hydrogenated overnight in a hydrogenatmosphere at standard pressure. The reaction solution was filteredthrough Celite, the filter residue was washed with methanol/water andthe combined filtrates were concentrated under reduced pressure. Theresidue was taken up in dichloromethane and then washed with a 1 Naqueous sodium carbonate solution. The organic phase was dried oversodium sulphate, filtered and concentrated under reduced pressure.Yield: 2.30 g (81% of theory, purity 82%)

LC-MS (Method 2B): R_(t)=0.80 and 0.81 min (cis/trans isomers); MS(ESIpos): m/z=284 [M+H]±.

Example 51A 3-Methyl 1-(4-nitrophenyl)5-[4-(1,1-difluoroethyl)phenyl]piperidine-1,3-dicarboxylate [racemiccis/trans isomer mixture]

A solution of 1.30 g (3.78 mmol, purity 82%) of the compound fromExample 50A in dichloromethane (44 ml) was admixed with 1.84 ml (1.34 g,13.2 mmol) of triethylamine and then admixed at 0° C. with 762 mg (3.78mmol) of 4-nitrophenyl chloroformate. The mixture was warmed to RT andstirred for 2 days. The reaction solution was washed with saturatedaqueous sodium hydrogencarbonate solution and water, and the organicphase was dried over magnesium sulphate, filtered and concentrated underreduced pressure. Yield: 1.93 g (92% of theory, purity 81%, 2:1 mixtureof methyl and ethyl ester)

LC-MS (Method 5B): R_(t)=2.58 min and 2.61 (methyl ester, cis/transisomers) and 2.68 and 2.70 (ethyl ester, cis/trans isomers); MS(ESIpos): m/z=278 [M+H]+(methyl ester) and 292 [M+H]+(ethyl ester).

Example 52A Methyl5-[4-(1,1-difluoroethyl)phenyl]-1-(thiomorpholin-4-ylcarbonyl)piperidine-3-carboxylate[racemic cis/trans isomer mixture]

A solution of 1.94 g (3.50 mmol, purity 81%) of the compound fromExample 51A in 1-methyl-2-pyrrolidone (18 ml) was admixed with 1.99 ml(2.17 g, 21.0 mmol) of thiomorpholine and 1.83 ml (1.36 g, 10.5 mmol) ofN,N-diisopropylethylamine and then heated in 3 portions in a single-modemicrowave (Emrys Optimizer) at 150° C. for 45 min For workup, thereaction solutions were combined and purified directly by means ofpreparative HPLC. Yield: 530 mg (34% of theory)

LC-MS (Method 5B): R_(t)=2.28 and 2.35 min (cis/trans isomers); MS(ESIpos): m/z=413 [M+H]⁺.

Example 53A5-[4-(1,1-Difluoroethyl)phenyl]-1-(thiomorpholin-4-ylcarbonyl)piperidine-3-carboxylicacid [racemic cis/trans isomer mixture]

1.44 g (12.8 mmol) of potassium tert-butoxide were added at RT to asolution of 528 mg (1.28 mmol) of the compound from Example 52A in 15 mlof methanol. The mixture was stirred at 60° C. overnight. For workup,the methanol was removed under reduced pressure, the residue was admixedwith water and the mixture was acidified (pH 1) with aqueous 1 Nhydrochloric acid solution. The mixture was extracted with ethylacetate, and the organic phase was dried with magnesium sulphate,filtered and concentrated under reduced pressure. Yield: 471 mg (91% oftheory, 2:1 cis/trans isomer mixture)

LC-MS (Method 5B): R_(t)=0.99 and 1.01 min; MS (ESIpos): m/z=399 [M+H]⁺.

Example 54A{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(1,1-difluoroethyl)phenyl]piperidin-1-yl}-(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 150 mg (0.376 mmol) of the compound fromExample 53A and 41.5 mg (0.414 mmol) ofN′-hydroxycyclopropanecarboximidamide were reacted Yield: 77.9 mg (44%of theory)

LC-MS (Method 2B): R_(t)=1.37 min; MS (ESIpos): m/z=463 [M+H]⁺.

Example 55A Methyl 5-[4-(2-hydroxyethyl)phenyl]nicotinate

According to General Method 3A, 6.00 g (29.8 mmol) of2-(4-bromophenyl)ethanol and 19.6 g (74.6 mmol) of methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)nicotinate were reacted.Yield: 6.12 g (74% of theory)

LC-MS (Method 2B): R_(t)=0.86 min; MS (ESIpos): m/z=258 [M+H]⁺.

Example 56A Methyl 5-[4-(2-hydroxyethyl)phenyl]piperidine-3-carboxylate[racemic cis/trans isomer mixture]

A solution of 5.40 g (19.6 mmol) of the compound from Example 55A inconcentrated acetic acid (124 ml) was admixed with 1.00 g ofpalladium/carbon (10% palladium) and 1.00 g of platinum(IV) oxide. Thiswas followed by hydrogenation under a hydrogen atmosphere at standardpressure for 6 h, then addition of another 1.00 g of palladium/carbon(10% palladium) and 1.00 g of platinum(IV) oxide, and hydrogenationunder a hydrogen atmosphere at standard pressure overnight. This wasfollowed by hydrogenation in a Parr apparatus under a 3 bar hydrogenatmosphere for a further 2 h. The reaction solution was filtered throughCelite, the filter residue was washed with methanol/water and thecombined filtrates were concentrated under reduced pressure. The residuewas codistilled repeatedly with toluene and then dried under highvacuum. Yield: 6.63 g (56% of theory, purity 44%)

LC-MS (Method 2B): R_(t)=0.36 min and 0.40 min (cis/trans isomers); MS(ESIpos): m/z=264 [M+H]⁺.

Example 57A

Methyl 1-acetyl-5-[4-(2-hydroxyethyl)phenyl]piperidine-3-carboxylate[racemic cis/trans isomer mixture]

A solution of 5.58 g (9.33 mmol, purity 44%) of the compound fromExample 56A in dichloromethane (80 ml) was admixed with 2.60 ml (1.89 g,18.7 mmol) of triethylamine and then cooled to 0° C. At thistemperature, 0.33 ml (0.37 g, 4.67 mmol) of acetyl chloride was addeddropwise and the mixture was stirred for 2 h. A further 0.13 ml (0.15 g,1.86 mmol) of acetyl chloride was added and the mixture was stirred for1 h. Subsequently, the reaction solution was washed with aqueous 1 Nhydrochloric acid, and the organic phase was dried over magnesiumsulphate, filtered and concentrated under reduced pressure. The residuewas purified by means of column chromatography (silica gel,dichloromethane/methanol 30:1), and the crude product obtained waspurified once more by means of preparative HPLC. Yield: 1.17 g (41% oftheory)

LC-MS (Method 5B): R_(t)=0.72 min and 0.74 min (cis/trans isomers); MS(ESIpos): m/z=306 [M+H]⁺.

Example 58A

Methyl 1-acetyl-5-[4-(2,2-difluoroethyl)phenyl]piperidine-3-carboxylate[racemic cis/trans isomer mixture]

A solution of 631 mg (2.05 mmol) of the compound from Example 57A indichloromethane (20.8 ml) was admixed with 1.45 ml (1.60 g, 20.5 mmol)of dimethyl sulphoxide and 1.78 ml (1.32 g, 10.2 mmol) ofN,N-diisopropylamine. Subsequently, 1.30 g (8.18 mmol) of sulphurtrioxide-pyridine complex were added at −20° C. and the mixture wasstirred overnight, in the course of which it was warmed slowly to RT.The reaction solution was diluted with dichloromethane, and the organicphase was washed with water, dried over magnesium sulphate, filtered andconcentrated under reduced pressure. The crude product (778 mg) wassubsequently initially charged in dichloromethane (5.2 ml) and admixeddropwise at RT with 0.50 ml (615 mg, 3.81 mmol) of diethylaminosulphurtrifluoride (DAST). The mixture was stirred at RT for 4 h and then thereaction was ended by cautiously adding 2 N aqueous sodium carbonatesolution. After phase separation, the organic phase was dried overmagnesium sulphate, filtered and concentrated under reduced pressure.The crude product was purified by means of preparative HPLC. Yield: 120mg (24% of theory, purity 61%, approx. 2:1 cis/trans isomer mixture)

LC-MS (Method 2B): R_(t)=1.07 min and 1.09 min (cis/trans isomers); MS(ESIpos): m/z=326 [M+H]⁺.

Example 59A1-Acetyl-5-[4-(2,2-difluoroethyl)phenyl]piperidine-3-carboxylic acid[racemic cis isomer]

410 mg (3.65 mmol) of potassium tert-butoxide were added at RT to asolution of 205 mg (0.365 mmol, purity 61%) of the compound from Example58A in methanol (6.9 ml). The mixture was stirred at 60° C. overnight.For workup, the methanol was removed under reduced pressure, the residuewas admixed with water and the mixture was acidified (pH 1) with aqueous1 N hydrochloric acid solution. The mixture was extracted with ethylacetate, and the organic phase was dried with magnesium sulphate,filtered and concentrated under reduced pressure. Yield: 153 mg (67% oftheory, purity 50%)

LC-MS (Method 5B): R_(t)=1.74 min; MS (ESIpos): m/z=312 [M+H]⁺.

Example 60A1-{3-[4-(2,2-Difluoroethyl)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}-ethanone[racemic cis isomer]

According to General Method 6A, 153 mg (0.270 mmol, purity 50%) of thecompound from Example 59A and 41.3 mg (0.297 mmol, purity 85%) ofN′-hydroxy-3-methoxypropanimidamide were reacted Yield: 46.6 mg (32% oftheory, purity 72%)

LC-MS (Method 2B): R_(t)=1.07 min; MS (ESIpos): m/z=394 [M+H]⁺.

Example 61A3-[4-(2,2-Difluoroethyl)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidine[racemic cis isomer]

A solution of 45.0 mg (0.083 mmol, purity 72%) of the compound fromExample 60A in ethanol (10.0 ml) was admixed with 69 μl (15 mg, 0.42mmol) of concentrated hydrochloric acid. Subsequently, the mixture wasstirred under reflux for 24 h, and the reaction solution was dilutedwith water and washed with diethyl ether. The aqueous phase wasalkalized and extracted with dichloromethane. The organic phase wasdried over magnesium sulphate, filtered and concentrated under reducedpressure. Yield: 36.3 mg (87% of theory, purity 70%)

LC-MS (Method 5B): R_(t)=0.71 min; MS (ESIpos): m/z=352 [M+H]⁺.

Example 62A 4-Nitrophenyl3-[4-(2,2-difluoroethyl)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidine-1-carboxylate[racemic cis isomer]

A solution of 36.3 mg (0.061 mmol, purity 70%) of the compound fromExample 61A in dichloromethane (2.0 ml) was admixed with 0.03 ml (21.6mg, 0.21 mmol) of triethylamine and then 12.3 mg (0.061 mmol) of4-nitrophenyl chloroformate were added at RT. The mixture was stirred atRT for 2 h and then the reaction solution was washed with saturatedaqueous sodium hydrogencarbonate solution and water, and the organicphase was dried over magnesium sulphate, filtered and concentrated underreduced pressure. Yield: 56.2 mg (91% of theory, purity 60%)

LC-MS (Method 4B): R_(t)=2.56 min; MS (ESIpos): m/z=517 [M+H]⁺.

Example 63A{3-[4-(2,2-Difluoroethyl)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(thiomorpholin-4-yl)methanone[racemic cis isomer]

A solution of 56.0 mg (0.065 mmol, purity 60%) of the compound fromExample 62A in 1-methyl-2-pyrrolidone (2.0 ml) was admixed with 37.0 pi(40.0 mg, 0.390 mmol) of thiomorpholine and 34.0 pi (25.0 mg, 0.195mmol) of N,N-diisopropylethylamine and then heated in a single-modemicrowave (Emrys Optimizer) at 150° C. for 30 min For workup, thereaction solutions were combined and purified directly by means ofpreparative HPLC. Yield: 14.7 mg (47% of theory)

LC-MS (Method 5B): R_(t)=1.09 min; MS (ESIpos): m/z=481 [M+H]⁺.

Example 64A N-Hydroxy-1-methoxycyclopropanecarboximidamide

100 mg (1.03 mmol) of 1-methoxycyclopropanecarboxamide [L. N. Owen, H.M. Babatunde Somade, J. Chem. Soc. 1947, 1030-1034] in tetrahydrofuran(22.7 ml) was admixed with 1.51 g (6.08 mmol) of methylN-(triethylammoniumsulphonyl)carbamate (Burgess reagent) and thenstirred at 60° C. for 1.5 h. The reaction mixture was admixed withdichloromethane and water, and the organic phase was dried overmagnesium sulphate, filtered and concentrated under reduced pressure(637 mg of crude product). 100 mg of the crude product were initiallycharged in ethanol (1.2 ml), admixed with 107 mg (1.55 mmol) ofhydroxylammonium chloride and 0.17 ml (125 mg, 1.24 mg) of triethylamineand then stirred under reflux overnight. The reaction solution wasconcentrated under reduced pressure, and the residue was admixed withsaturated aqueous sodium chloride solution and then extracted withdichloromethane. The organic phase was dried over magnesium sulphate,filtered and concentrated under reduced pressure. The residue wassubsequently stirred with ethyl acetate, the insoluble salts werefiltered off and the filtrate was concentrated under reduced pressure.Yield: 32.3 mg (23% of theory)

¹H NMR (400 MHz, DMSO-d₆): δ=9.09 (br. s., 1H), 5.39 (br. s., 2H), 3.15(s, 3H), 0.81 (d, 4H).

Example 65A{3-[3-(1-Methoxycyclopropyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(thiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 6A, 93.1 mg (0.224 mmol) of the compoundfrom Example 45A and 32.0 mg (0.246 mmol) ofN′-hydroxy-1-methoxycyclopropanecarboximidamide from Example 64A werereacted. Yield: 31.1 mg (27% of theory)

LC-MS (Method 5B): R_(t)=1.22 min; MS (ESIpos): m/z=511 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): δ=7.33 (s, 4H), 3.94 (d, 1H), 3.69-3.52 (m,3H), 3.48-3.42 (m, 4H), 3.41-3.34 (m, 4H), 3.06-2.85 (m, 3H), 2.63-2.57(m, 4H), 2.32-2.25 (m, 1H), 2.02-1.88 (m, 1H), 1.34-1.28 (m, 2H),1.19-1.11 (m, 2H).

Example 66A 4-Nitrophenyl thiomorpholine-4-carboxylate 1,1-dioxide

17.0 g (99.2 mmol) of thiomorpholine 1,1-dioxide hydrochloride wereinitially charged in 100 ml of dichloromethane and, while cooling withan ice bath, admixed with 20.7 ml (15.1 g, 148.8 mmol) of triethylamine10.0 g (49.6 mmol) of 4-nitrophenyl chloroformate were added inportions. The reaction mixture was stirred at RT for 30 minutes, admixedwith water and ethyl acetate and then filtered. The residue was driedunder high vacuum. Yield: 12.4 g (83% of theory)

LC-MS (Method 5B): R_(t)=0.75 min; MS (ESIpos): m/z=301 [M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆): 6=8.34-8.28 (m, 2H), 7.55-7.50 (m, 2H), 4.01(br. s., 2H), 3.87 (br. s., 2H), 3.37 (br. s., 2H), 3.28 (br. s., 2H).

Working Examples General Method 1: Sulphoxide Formation

A solution of the appropriate thioether (1.0 eq) in dichloromethane(approx. 20-40 ml/mmol) is admixed at RT with 50%meta-chloroperoxybenzoic acid (0.9-1.0 eq.). The reaction mixture isstirred at room temperature for 30 min The solvent is removed underreduced pressure and the residue is purified by means of preparativeHPLC.

General Method 2: Sulphone Formation

A solution of the appropriate thioether (1.0 eq) in dichloromethane(approx. 20-40 ml/mmol) is admixed at RT with 50%meta-chloroperoxybenzoic acid (2.5 eq.). The reaction mixture is stirredat room temperature for 30 min. The solvent is removed under reducedpressure and the residue is purified by means of preparative HPLC.

Example 1{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}-(oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 100 mg (0.200 mmol) of the compound fromExample 26A were reacted. Yield: 90 mg (87% of theory)

LC-MS (Method 5B): R_(t)=0.97 min; MS (ESIpos): m/z=517 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.48 (d, 2H), 7.33 (d, 3H), 3.99 (d, 1H),3.69-3.66 (m, 3H), 3.65-3.58 (m, 4H), 3.57-3.48 (m, 3H), 3.23 (s, 3H),3.09-2.88 (m, 7H), 2.75-2.66 (m, 3H), 2.35-2.28 (m, 1H), 2.00 (m, 1H).

Example 2{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 100 mg (0.207 mmol) of the compound fromExample 27A were reacted. Yield: 95 mg (91% of theory)

LC-MS (Method 5B): R_(t)=1.05 min; MS (ESIpos): m/z=499 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.47 (d, 2H), 7.33 (d, 2H), 3.95 (d, 1H),3.67-3.48 (m, 5H), 3.05-2.85 (m, 5H), 2.75-2.65 (m, 2H), 2.28 (d, 1H),2.14-2.08 (m, 1H), 1.94 (q, 1H), 1.09-1.01 (m, 2H), 0.92-0.84 (m, 2H).

Example 3[3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-(4-ethylphenyl)piperidin-1-yl](1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 55 mg (0.130 mmol) of the compound fromExample 9A were reacted. Yield: 43 mg (75% of theory)

LC-MS (Method 5B): R_(t)=1.06 min; MS (ESIpos): m/z=443 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.24 (d, 2H), 7.17 (d, 2H), 3.96 (d, 2H),3.67-3.46 (m, 5H), 3.045-2.85 (m, 5H), 2.74-2.66 (m, 2H), 2.57 (q, 2H),2.25 (d, 1H), 2.14-2.06 (m, 1H), 1.91 (q, 1H), 1.16 (t, 3H), 1.08-1.02(m, 2H), 0.91-0.86 (m, 2H.)

Example 4{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-(4-ethylphenyl)piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 50 mg (0.109 mmol) of the compound fromExample 11A were reacted. Yield: 17 mg (32% of theory)

LC-MS (Method 5B): R_(t)=1.01 min; MS (ESIpos): m/z=475 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.17 (d, 2H), 3.98 (d, 2H),3.71 (t, 2H), 3.67-3.47 (m, 4H), 3.43 (q, 1H), 3.07-2.89 (m, 3H),2.75-2.66 (m, 3H), 2.57 (q, 3H), 1.95 (q, 1H), 1.24 (br s, 1H), 1.16 (t,3H), 1.07 (t, 3H).

Example 5{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-(4-ethylphenyl)piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of the racemate from Example 4 according to Method6D gave 37.7 mg of the title compound from Example 5 and 20.0 mg of thetitle compound from Example 6.

LC-MS (Method 5B): R_(t)=1.01 min; MS (ESIpos): m/z=475 [M+H]⁺;

HPLC (Method 1E): R_(t)=6.48 min, >99.5% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.17 (d, 2H), 3.98 (d, 2H),3.71 (t, 2H), 3.67-3.47 (m, 4H), 3.43 (q, 1H), 3.07-2.89 (m, 3H),2.75-2.66 (m, 3H), 2.57 (q, 3H), 1.95 (q, 1H), 1.24 (br s, 1H), 1.16 (t,3H), 1.07 (t, 3H).

Example 6{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-(4-ethylphenyl)piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of the racemate from Example 4 according to Method6D gave 37.7 mg of the title compound from Example 5 and 20.0 mg of thetitle compound from Example 6.

LC-MS (Method 5B): R_(t)=1.01 min; MS (ESIpos): m/z=475 [M+H]⁺;

HPLC (Method 1E): R_(t)=7.27 min, >99.5% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.17 (d, 2H), 3.98 (d, 2H),3.71 (t, 2H), 3.67-3.47 (m, 4H), 3.43 (q, 1H), 3.07-2.89 (m, 3H),2.75-2.66 (m, 3H), 2.57 (q, 3H), 1.95 (q, 1H), 1.24 (br s, 1H), 1.16 (t,3H), 1.07 (t, 3H).

Example 7{3-[3-(2-Hydroxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 80 mg (0.170 mmol) of the compound fromExample 20A were reacted. Yield: 77 mg (91% of theory)

LC-MS (Method 5B): R_(t)=0.85 min; MS (ESIpos): m/z=487 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.77 (t, 1H),3.99 (d, 1H), 3.74 (q, 2H), 3.68-3.59 (m, 3H), 3.57-3.48 (m, 2H),3.48-3.38 (m, 1H), 3.12-3.01 (m, 3H), 2.98-2.86 (m 2H), 2.85-2.80 (m,2H), 1.55 (q, 1H).

Example 8(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 150 mg (0.300 mmol) of the compound fromExample 26A were reacted. Enantiomer separation of the racemateaccording to Method 1D gave 65.0 mg of the title compound from Example 8and 72.0 mg of the title compound from Example 9.

LC-MS (Method 5B): R_(t)=1.04 min; MS (ESIpos): m/z=533 [M+H]⁺;

HPLC (Method 2E): R_(t)=15.64 min, >99.5% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.48 (d, 2H), 7.33 (d, 2H), 4.03 (d, 1H),3.70-3.58 (m, 7H), 3.45-3.35 (m, 1H), 3.23 (s, 3H), 3.21-3.15 (m, 4H),3.12-2.90 (m, 5H), 2.33 (d, 1H), 1.97 (q, 1H).

Example 9(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 150 mg (0.300 mmol) of the compound fromExample 26A were reacted. Enantiomer separation of the racemateaccording to Method 1D gave 65.0 mg of the title compound from Example 8and 72.0 mg of the title compound from Example 9.

LC-MS (Method 5B): R_(t)=1.04 min; MS (ESIpos): m/z=533 [M+H]⁺;

HPLC (Method 2E): R_(t)=42.42 min, >99.5% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.48 (d, 2H), 7.33 (d, 2H), 4.03 (d, 1H),3.70-3.58 (m, 7H), 3.45-3.35 (m, 1H), 3.23 (s, 3H), 3.21-3.15 (m, 4H),3.12-2.90 (m, 5H), 2.33 (d, 1H), 1.97 (q, 1H).

Example 10{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

136 mg (0.281 mmol) of{3-(3-cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethoxy)phenyl]-piperidin-1-yl}(thiomorpholin-4-yl)methanone[racemic cis isomer] (Example 27A) in dichloromethane (11.6 ml) wereadmixed at RT with 243 mg (0.703 mmol) of meta-chloroperbenzoic acid andthen stirred for 30 min The reaction solution was concentrated underreduced pressure, and the residue was taken up in acetonitrile andpurified by means of preparative HPLC. Enantiomer separation of 136 mgof the racemate according to Method 2D gave 61.7 mg of the titlecompound from Example 10 and 59.6 mg of the title compound from Example11.

LC-MS (Method 5B): R_(t)=1.12 min; MS (ESIpos): m/z=515 [M+H]⁺;

HPLC (Method 3E): R_(t)=4.26 min, >99.05% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.47 (d, 2H), 7.33 (d, 2H), 3.99 (d, 1H),3.67-3.56 (m, 5H), 3.40-3.33 (m, 1H), 3.20-3.14 (m, 4H), 3.08-2.96 (m,3H), 2.28 (d, 1H), 2.14-2.06 (m, 1H), 1.94 (q, 1H), 1.08-1.03 (m, 2H),0.91-0.86 (m, 2H).

Example 11{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

136 mg (0.281 mmol) of{3-(3-cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethoxy)phenyl]-piperidin-1-yl}(thiomorpholin-4-yl)methanone[racemic cis isomer] (Example 27A) in dichloromethane (11.6 ml) wereadmixed at RT with 243 mg (0.703 mmol) of meta-chloroperbenzoic acid andthen stirred for 30 min. The reaction solution was concentrated underreduced pressure, and the residue was taken up in acetonitrile andpurified by means of preparative HPLC. Enantiomer separation of 136 mgof the racemate according to Method 2D gave 61.7 mg of the titlecompound from Example 10 and 59.6 mg of the title compound from Example11.

LC-MS (Method 5B): R_(t)=1.12 min; MS (ESIpos): m/z=515 [M+H]⁺;

HPLC (Method 3E): R_(t)=5.68 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.47 (d, 2H), 7.33 (d, 2H), 3.99 (d, 1H),3.67-3.56 (m, 5H), 3.40-3.33 (m, 1H), 3.20-3.14 (m, 4H), 3.08-2.96 (m,3H), 2.28 (d, 1H), 2.14-2.06 (m, 1H), 1.94 (q, 1H), 1.08-1.03 (m, 2H),0.91-0.86 (m, 2H).

Example 12(1,1-Dioxidothiomorpholin-4-yl){3-(4-ethylphenyl)-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 77.0 mg (0.173 mmol) of the compound fromExample 8A were reacted. Enantiomer separation of 74.9 mg of theracemate according to Method 3D gave 36.0 mg of the title compound fromExample 12 and 35.0 mg of the title compound from Example 13.

LC-MS (Method 2B): R_(t)=1.17 min; MS (ESIpos): m/z=477 [M+H]⁺;

HPLC (Method 4E): R_(t)=5.49 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.17 (d, 2H), 4.03 (d, 1H),3.73-3.56 (m, 7H), 3.46-3.35 (m, 1H), 3.23 (s, 3H), 3.17 (br s, 4H),3.06 (t, 1H), 3.01-2.83 (m, 4H), 2.58 (d, 3H), 2.30 (d, 1H), 1.95 (q,1H), 1.16 (t, 3H).

Example 13(1,1-Dioxidothiomorpholin-4-yl){3-(4-ethylphenyl)-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 77.0 mg (0.173 mmol) of the compound fromExample 8A were reacted. Enantiomer separation of 74.9 mg of theracemate according to Method 3D gave 36.0 mg of the title compound fromExample 12 and 35.0 mg of the title compound from Example 13.

LC-MS (Method 2B): R_(t)=1.17 min; MS (ESIpos): m/z=477 [M+H]⁺;

HPLC (Method 4E): R_(t)=12.07 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.17 (d, 2H), 4.03 (d, 1H),3.73-3.56 (m, 7H), 3.46-3.35 (m, 1H), 3.23 (s, 3H), 3.17 (br s, 4H),3.06 (t, 1H), 3.01-2.83 (m, 4H), 2.58 (d, 3H), 2.30 (d, 1H), 1.95 (q,1H), 1.16 (t, 3H).

Example 14[3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-(4-ethylphenyl)piperidin-1-yl](1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

According to General Method 2, 55 mg (0.130 mmol) of the compound fromExample 9A were reacted. Enantiomer separation of 53.3 mg of theracemate according to Method 4D gave 23.0 mg of the title compound fromExample 14 and 23.0 mg of the title compound from Example 15.

LC-MS (Method 2B): R_(t)=1.30 min; MS (ESIpos): m/z=459 [M+H]⁺;

HPLC (Method 5E): R_(t)=8.89 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.16 (d, 2H), 3.99 (d, 1H),3.67-3.55 (m, 5H), 3.39-3.32 (m, 1H), 3.17 (br s, 4H), 3.07-2.91 (m,2H), 2.91-2.81 (m, 1H), 2.62-2.55 (m, 2H), 2.26 (d, 1H), 2.16-2.08 (m,1H), 1.91 (q, 1H), 1.16 (t, 3H), 1.10-1.02 (m, 2H), 0.92-0.85 (m, 2H).

Example 15[3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-(4-ethylphenyl)piperidin-1-yl](1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

According to General Method 2, 55.5 mg (0.130 mmol) of the compound fromExample 9A were reacted. Enantiomer separation of 53.3 mg of theracemate according to Method 4D gave 23.0 mg of the title compound fromExample 14 and 23.0 mg of the title compound from Example 15.

LC-MS (Method 2B): R_(t)=1.27 min; MS (ESIpos): m/z=459 [M+H]⁺;

HPLC (Method 5E): R_(t)=12.06 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (d, 2H), 7.16 (d, 2H), 3.99 (d, 1H),3.67-3.55 (m, 5H), 3.39-3.32 (m, 1H), 3.17 (br s, 4H), 3.07-2.91 (m,2H), 2.91-2.81 (m, 1H), 2.62-2.55 (m, 2H), 2.26 (d, 1H), 2.16-2.08 (m,1H), 1.91 (q, 1H), 1.16 (t, 3H), 1.10-1.02 (m, 2H), 0.92-0.85 (m, 2H).

Example 16{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

According to General Method 2, 269 mg (0.578 mmol) of the compound fromExample 17A were reacted. Enantiomer separation of 292 mg of theracemate according to Method 1D gave 57.8 mg of the title compound fromExample 16 and 99.7 mg of the title compound from Example 17.

LC-MS (Method 2B): R_(t)=1.26 min; MS (ESIpos): m/z=499 [M+H]⁺;

HPLC (Method 1E): R_(t)=10.53 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.57 (d, 2H), 4.00 (d, 1H),3.67 (d, 1H), 3.61 (br s, 4H), 3.44-3.33 (m, 1H), 3.17 (br s, 4H),3.12-2.98 (m, 3H), 2.31 (d, 1H), 2.11 (dt, 1H), 1.98 (q, 1H), 1.12-1.00(m, 2H), 0.95-0.84 (m, 2H).

Example 17{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

According to General Method 2, 269 mg (0.578 mmol) of the compound fromExample 17A were reacted. Enantiomer separation of 292 mg of theracemate according to Method 1D gave 57.8 mg of the title compound fromExample 16 and 99.7 mg of the title compound from Example 17.

LC-MS (Method 2B): R_(t)=1.26 min; MS (ESIpos): m/z=499 [M+H]⁺;

HPLC (Method 1E): R_(t)=16.04 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.57 (d, 2H), 4.00 (d, 1H),3.67 (d, 1H), 3.61 (br s, 4H), 3.44-3.33 (m, 1H), 3.17 (br s, 4H),3.12-2.98 (m, 3H), 2.31 (d, 1H), 2.11 (dt, 1H), 1.98 (q, 1H), 1.12-1.00(m, 2H), 0.95-0.84 (m, 2H).

Example 18 (1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 317 mg (0.616 mmol) of the compound fromExample 28A were reacted. Enantiomer separation of 294 mg of theracemate according to Method 2D gave 132 mg of the title compound fromExample 18 and 129 mg of the title compound from Example 19.

LC-MS (Method 6B): R_(t)=2.34 min; MS (ESIpos): m/z=547 [M+H]⁺;

HPLC (Method 3E): R_(t)=4.60 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.48 (d, 2H), 7.33 (d, 2H), 4.03 (d, 1H),3.71 (t, 2H), 3.68-3.56 (m, 5H), 3.43 (q, 3H), 3.17 (br s, 4H), 3.07 (t,1H), 3.01 (d, 2H), 2.93 (t, 2H), 2.32 (d, 1H), 2.05-1.91 (m, 1H), 1.07(t, 3H).

Example 19(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 317 mg (0.616 mmol) of the compound fromExample 28A were reacted. Enantiomer separation of 294 mg of theracemate according to Method 2D gave 132 mg of the title compound fromExample 18 and 129 mg of the title compound from Example 19.

LC-MS (Method 6B): R_(t)=2.34 min; MS (ESIpos): m/z=547 [M+H]⁺;

HPLC (Method 3E): R_(t)=11.53 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.48 (d, 2H), 7.33 (d, 2H), 4.03 (d, 1H),3.71 (t, 2H), 3.68-3.56 (m, 5H), 3.43 (q, 3H), 3.17 (br s, 4H), 3.07 (t,1H), 3.01 (d, 2H), 2.93 (t, 2H), 2.32 (d, 1H), 2.05-1.91 (m, 1H), 1.07(t, 3H).

Example 20{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 50.0 mg (0.107 mmol) of the compound fromExample 17A were reacted. Yield: 4.3 mg (8% of theory)

LC-MS (Method 5B): R_(t)=1.05 min; MS (ESIpos): m/z=483 [M+H]⁺.

Example 21{3-[3-(2-Hydroxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}-(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 100.0 mg (0.206 mmol) of the compoundfrom Example 29A were reacted. Yield: 105.2 mg (99% of theory)

LC-MS (Method 5B): R_(t)=0.89 min; MS (ESIpos): m/z=503 [M+H]⁺.

Example 22{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 50.0 mg (0.100 mmol) of the compound fromExample 19A were reacted. Yield: 50.2 mg (92% of theory)

LC-MS (Method 5B): R_(t)=1.02 min; MS (ESIpos): m/z=515 [M+H]⁺;

Example 23{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 50.2 mg of the racemate from Example 22according to Method 1D gave 25.5 mg of the title compound from Example23 and 22.4 mg of the title compound from Example 24.

LC-MS (Method 2B): R_(t)=1.14 min; MS (ESIpos): m/z=515 [M+H]⁺;

HPLC (Method 2E): R_(t)=7.51 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.00 (d, 1H),3.71 (t, 2H), 3.63 (d, 3H), 3.57-3.48 (m, 2H), 3.43 (q, 3H), 3.14-3.00(m, 3H), 2.93 (t, 4H), 2.77-2.65 (m, 2H), 2.35 (d, 1H), 2.10-1.95 (m,1H), 1.06 (m, 3H).

Example 24{3-[3-(2-Ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 50.2 mg of the racemate from Example 22according to Method 1D gave 25.5 mg of the title compound from Example23 and 22.4 mg of the title compound from Example 24.

LC-MS (Method 2B): R_(t)=1.14 min; MS (ESIpos): m/z=515 [M+H]⁺;

HPLC (Method 2E): R_(t)=13.93 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.00 (d, 1H),3.71 (t, 2H), 3.63 (d, 3H), 3.57-3.48 (m, 2H), 3.43 (q, 3H), 3.14-3.00(m, 3H), 2.93 (t, 4H), 2.77-2.65 (m, 2H), 2.35 (d, 1H), 2.10-1.95 (m,1H), 1.06 (m, 3H).

Example 25{3-(4-Ethylphenyl)-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(1-oxidothio-morpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 77.0 mg (0.173 mmol) of the compound fromExample 8A were reacted. Yield: 63.2 mg (79% of theory)

LC-MS (Method 5B): R_(t)=0.97 min; MS (ESIpos): m/z=461 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (m, 2H), 7.17 (m, 2H), 3.99 (d, 1H),3.72-3.46 (m, 7H), 3.46-3.34 (m, 2H), 3.09-2.98 (m, 1H), 2.97-2.83 (m,6H), 2.65 (br s, 1H), 2.76-2.64 (m, 2H), 2.58 (d, 3H), 2.30 (d, 1H),1.95 (q, 1H), 1.16 (t, 3H); one proton hidden.

Example 26{3-(4-Ethylphenyl)-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(1-oxidothio-morpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 63.2 mg of the racemate from Example 25according to Method 6D gave 17.8 mg of the title compound from Example26 and 18.7 mg of the title compound from Example 27.

LC-MS (Method 5B): R_(t)=0.97 min; MS (ESIpos): m/z=461 [M+H]⁺;

HPLC (Method 2E): R_(t)=6.48 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (m, 2H), 7.17 (m, 2H), 3.99 (d, 1H),3.72-3.46 (m, 7H), 3.46-3.34 (m, 2H), 3.09-2.98 (m, 1H), 2.97-2.83 (m,6H), 2.65 (br s, 1H), 2.76-2.64 (m, 2H), 2.58 (d, 3H), 2.30 (d, 1H),1.95 (q, 1H), 1.16 (t, 3H); one proton hidden.

Example 27{3-(4-Ethylphenyl)-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(1-oxidothio-morpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 63.2 mg of the racemate from Example 25according to Method 6D gave 17.8 mg of the title compound from Example26 and 18.7 mg of the title compound from Example 27.

LC-MS (Method 5B): R_(t)=0.97 min; MS (ESIpos): m/z=461 [M+H]⁺;

HPLC (Method 2E): R_(t)=7.97 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.23 (m, 2H), 7.17 (m, 2H), 3.99 (d, 1H),3.72-3.46 (m, 7H), 3.46-3.34 (m, 2H), 3.09-2.98 (m, 1H), 2.97-2.83 (m,6H), 2.65 (br s, 1H), 2.76-2.64 (m, 2H), 2.58 (d, 3H), 2.30 (d, 1H),1.95 (q, 1H), 1.16 (t, 3H); one proton hidden.

Example 28 (1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 269 mg (0.556 mmol) of the compound fromExample 18A were reacted. Enantiomer separation of the racemateaccording to Method 5D gave 126 mg of the title compound from Example 28and 122 mg of the title compound from Example 29.

LC-MS (Method 6B): R_(t)=2.18 min; MS (ESIpos): m/z=517 [M+H]⁺;

HPLC (Method 2E): R_(t)=4.98 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.03 (d, 1H),3.68 (t, 3H), 3.62 (br s, 4H), 3.49-3.38 (m, 1H), 3.23 (s, 3H), 3.18 (brs, 4H), 3.14-3.02 (m, 3H), 2.94 (t, 2H), 2.35 (d, 1H), 2.02 (d, 1H).

Example 29(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 269 mg (0.556 mmol) of the compound fromExample 18A were reacted. Enantiomer separation of the racemateaccording to Method 5D gave 126 mg of the title compound from Example 28and 122 mg of the title compound from Example 29.

LC-MS (Method 6B): R_(t)=2.18 min; MS (ESIpos): m/z=517 [M+H]⁺;

HPLC (Method 2E): R_(t)=15.96 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.03 (d, 1H),3.68 (t, 3H), 3.62 (br s, 4H), 3.49-3.38 (m, 1H), 3.23 (s, 3H), 3.18 (brs, 4H), 3.14-3.02 (m, 3H), 2.94 (t, 2H), 2.35 (d, 1H), 2.02 (d, 1H).

Example 30 (1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 259 mg (0.519 mmol) of the compound fromExample 19A were reacted. Enantiomer separation of 252 mg of theracemate according to Method 2D gave 104 mg of the title compound fromExample 30 and 91.0 mg of the title compound from Example 31.

LC-MS (Method 6B): R_(t)=2.29 min; MS (ESIpos): m/z=531 [M+H]⁺;

HPLC (Method 3E): R_(t)=4.92 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.71 (d, 2H), 7.58 (d, 2H), 4.03 (d, 1H),3.75-3.68 (m, 3H), 3.62 (br s, 4H), 3.43 (q, 3H), 3.18 (br s, 4H),3.14-3.01 (m, 3H), 2.93 (t, 2H), 2.35 (d, 1H), 2.12-1.95 (m, 1H), 1.07(t, 3H).

Example 31(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 259 mg (0.519 mmol) of the compound fromExample 19A were reacted. Enantiomer separation of 252 mg of theracemate according to Method 2D gave 104 mg of the title compound fromExample 30 and 91.0 mg of the title compound from Example 31.

LC-MS (Method 6B): R_(t)=2.29 min; MS (ESIpos): m/z=531 [M+H]⁺;

HPLC (Method 3E): R_(t)=13.63 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.71 (d, 2H), 7.58 (d, 2H), 4.03 (d, 1H),3.75-3.68 (m, 3H), 3.62 (br s, 4H), 3.43 (q, 3H), 3.18 (br s, 4H),3.14-3.01 (m, 3H), 2.93 (t, 2H), 2.35 (d, 1H), 2.12-1.95 (m, 1H), 1.07(t, 3H).

Example 32(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-hydroxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 100 mg (0.213 mmol) of the compound fromExample 20A were reacted. Enantiomer separation of 97.4 mg of theracemate according to Method 2D gave 33.9 mg of the title compound fromExample 32 and 35.0 mg of the title compound from Example 33.

LC-MS (Method 5B): R_(t)=0.91 min; MS (ESIpos): m/z=503 [M+H]⁺;

HPLC (Method 3E): R_(t)=4.75 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.71 (d, 2H), 7.58 (d, 2H), 4.77 (t, 1H),4.03 (d, 1H), 3.74 (q, 2H), 3.68 (d, 1H), 3.62 (br s, 4H), 3.47-3.37 (m,1H), 3.18 (br s, 4H), 3.13-3.00 (m, 3H), 2.82 (t, 2-H), 2.35 (d, 1H),2.10-1.94 (m, 1H).

Example 33(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-hydroxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 100 mg (0.213 mmol) of the compound fromExample 20A were reacted. Enantiomer separation of 97.4 mg of theracemate according to Method 2D gave 33.9 mg of the title compound fromExample 32 and 35.0 mg of the title compound from Example 33.

LC-MS (Method 5B): R_(t)=0.91 min; MS (ESIpos): m/z=503 [M+H]⁺;

HPLC (Method 3E): R_(t)=8.97 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.71 (d, 2H), 7.58 (d, 2H), 4.77 (t, 1H),4.03 (d, 1H), 3.74 (q, 2H), 3.68 (d, 1H), 3.62 (br s, 4H), 3.47-3.37 (m,1H), 3.18 (br s, 4H), 3.13-3.00 (m, 3H), 2.82 (t, 2-H), 2.35 (d, 1H),2.10-1.94 (m, 1H).

Example 34(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-(4-ethylphenyl)-piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 303 mg (0.661 mmol) of the compound fromExample 11A were reacted. Enantiomer separation of 297 mg of theracemate according to Method 2D gave 139 mg of the title compound fromExample 34 and 117 mg of the title compound from Example 35.

LC-MS (Method 2B): R_(t)=1.24 min; MS (ESIpos): m/z=491 [M+H]⁺;

HPLC (Method 3E): R_(t)=4.81 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.24 (m, 2H), 7.17 (m, 2H), 4.03 (d, 1H),3.71 (t, 2H), 3.67-3.57 (m, 5H), 3.49-3.35 (m, 3H), 3.17 (br s, 4H),3.06 (t, 1H), 2.98-2.86 (m, 3H), 2.62-2.55 (m, 3H), 2.30 (d, 2H), 1.95(q, 1H), 1.16 (t, 3H), 1.07 (t, 3H).

Example 35(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-ethoxyethyl)-1,2,4-oxadiazol-5-yl]-5-(4-ethylphenyl)-piperidin-1-yl}methanone[enantiomerically pure cis isomer]

According to General Method 2, 303 mg (0.661 mmol) of the compound fromExample 11A were reacted. Enantiomer separation of 297 mg of theracemate according to Method 2D gave 139 mg of the title compound fromExample 34 and 117 mg of the title compound from Example 35.

LC-MS (Method 2B): R_(t)=1.24 min; MS (ESIpos): m/z=491 [M+H]⁺;

HPLC (Method 3E): R_(t)=6.80 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.24 (m, 2H), 7.17 (m, 2H), 4.03 (d, 1H),3.71 (t, 2H), 3.67-3.57 (m, 5H), 3.49-3.35 (m, 3H), 3.17 (br s, 4H),3.06 (t, 1H), 2.98-2.86 (m, 3H), 2.62-2.55 (m, 3H), 2.30 (d, 2H), 1.95(q, 1H), 1.16 (t, 3H), 1.07 (t, 3H).

Example 36{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

According to General Method 1, 196 mg (0.405 mmol) of the compound fromExample 18A were reacted. Yield: 194 mg (96% of theory)

LC-MS (Method 2B): R_(t)=1.08 min; MS (ESIpos): m/z=501 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.00 (d, 1H),3.73-3.58 (m, 5H), 3.57-3.48 (m, 2H), 3.48-3.39 (m, 1H), 3.13-2.99 (m,3H), 2.97-2.84 (m, 4H), 2.77-2.65 (m, 2H), 2.35 (d, 1H), 2.03 (q, 1H).

Example 37{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 194 mg of the racemate from Example 36according to Method 1D gave 81.1 mg of the title compound from Example37 and 78.5 mg of the title compound from Example 38.

LC-MS (Method 2B): R_(t)=1.08 min; MS (ESIpos): m/z=501 [M+H]⁺;

HPLC (Method 1E): R_(t)=8.45 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.00 (d, 1H),3.73-3.58 (m, 5H), 3.57-3.48 (m, 2H), 3.48-3.39 (m, 1H), 3.13-2.99 (m,3H), 2.97-2.84 (m, 4H), 2.77-2.65 (m, 2H), 2.35 (d, 1H), 2.03 (q, 1H).

Example 38{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(trifluoromethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 194 mg of the racemate from Example 36according to Method 1D gave 81.1 mg of the title compound from Example37 and 78.5 mg of the title compound from Example 38.

LC-MS (Method 2B): R_(t)=1.08 min; MS (ESIpos): m/z=501 [M+H]⁺;

HPLC (Method 1E): R_(t)=18.94 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.70 (d, 2H), 7.58 (d, 2H), 4.00 (d, 1H),3.73-3.58 (m, 5H), 3.57-3.48 (m, 2H), 3.48-3.39 (m, 1H), 3.13-2.99 (m,3H), 2.97-2.84 (m, 4H), 2.77-2.65 (m, 2H), 2.35 (d, 1H), 2.03 (q, 1H).

Example 39{3-[4-(2,2-Difluoroethyl)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[racemic cis isomer]

14.7 mg (0.031 mmol) of the compound from Example 63A were reactedaccording to General Method 2 with 26.3 mg (0.076 mmol) ofmeta-chloroperbenzoic acid. Yield: 9.5 mg (60% of theory)

¹H NMR (400 MHz, DMSO-d₆): δ=7.33 (d, 2H), 7.26 (m, 2H), 6.23 (tt, 1H),4.03 (d, 1H), 3.71-3.56 (m, 7H), 3.46-3.36 (m, 1H), 3.23 (s, 3H),3.21-3.13 (m, 5H), 3.12-2.87 (m, 6H), 2.31 (d, 1H), 1.97 (q, 1H).

Example 40{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

78.0 mg (0.162 mmol) of the compound from Example 46A were reactedaccording to General Method 1 with 50.4 mg (0.146 mmol) ofmeta-chloroperbenzoic acid. Yield: 76.2 mg (88% of theory)

LC-MS (Method 5B): R_(t)=1.02 min; MS (ESIpos): m/z=497 [M+H]⁺.

Example 41{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 76.2 mg of the racemate from Example 40according to Method 7D gave 29.1 mg of the title compound from Example41 (Enantiomer 1) and 28.9 mg of the title compound from Example 42(Enantiomer 2).

LC-MS (Method 7B): R_(t)=2.18 min; MS (ESIpos): m/z=497 [M+H]⁺;

HPLC (Method 6E): R_(t)=13.2 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.37-7.29 (m, 4H), 3.96 (d, 1H), 3.68-3.47(m, 7H), 3.41-3.33 (m, 1H), 3.07-2.85 (m, 5H), 2.74-2.66 (m, 2H), 2.28(d, 1H), 2.16-2.08 (m, 1H), 1.93 (q, 1H), 1.08-1.02 (m, 2H), 0.92-0.85(m, 2H).

Example 42{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 76.2 mg of the racemate from Example 40according to Method 7D gave 29.1 mg of the title compound from Example41 (Enantiomer 1) and 28.9 mg of the title compound from Example 42(Enantiomer 2).

LC-MS (Method 7B): R_(t)=2.18 min; MS (ESIpos): m/z=497 [M+H]⁺;

HPLC (Method 6E): R_(t)=16.4 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.37-7.29 (m, 4H), 3.96 (d, 1H), 3.68-3.47(m, 7H), 3.41-3.33 (m, 1H), 3.07-2.85 (m, 5H), 2.74-2.66 (m, 2H), 2.28(d, 1H), 2.16-2.08 (m, 1H), 1.93 (q, 1H), 1.08-1.02 (m, 2H), 0.92-0.85(m, 2H).

Example 43{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[racemic cis isomer]

78.0 mg (0.162 mmol) of the compound from Example 46A were reactedaccording to General Method 2 with 140 mg (0.146 mmol) ofmeta-chloroperbenzoic acid. Yield: 87.5 mg (100% of theory)

LC-MS (Method 2B): R_(t)=1.25 min; MS (ESIpos): m/z=513 [M+H]⁺.

Example 44{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 87.5 mg of the racemate from Example 43according to Method 8D gave 29.1 mg of the title compound from Example44 (Enantiomer 1) and 30.7 mg of the title compound from Example 45(Enantiomer 2).

LC-MS (Method 7B): R_(t)=2.34 min; MS (ESIpos): m/z=513 [M+H]⁺;

HPLC (Method 7E): R_(t)=9.86 min, 99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.37-7.29 (m, 4H), 4.00 (d, 1H), 3.67-3.56(m, 7H), 3.17 (br. s., 4H), 3.07-2.87 (m, 3H), 2.28 (d, 1H), 2.16-2.08(m, 1H), 1.93 (q, 1H), 1.09-1.02 (m, 2H), 0.92-0.85 (m, 3H), one protonhidden.

Example 45{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 87.5 mg of the racemate from Example 43according to Method 8D gave 29.1 mg of the title compound from Example44 (Enantiomer 1) and 30.7 mg of the title compound from Example 45(Enantiomer 2).

LC-MS (Method 7B): R_(t)=2.34 min; MS (ESIpos): m/z=513 [M+H]⁺;

HPLC (Method 7E): R_(t)=10.9 min, 97.5% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.37-7.29 (m, 4H), 4.00 (d, 1H), 3.67-3.56(m, 7H), 3.17 (br. s., 4H), 3.07-2.87 (m, 3H), 2.28 (d, 1H), 2.16-2.08(m, 1H), 1.93 (q, 1H), 1.09-1.02 (m, 2H), 0.92-0.85 (m, 3H), one protonhidden.

Example 46{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

113 mg (0.227 mmol) of the compound from Example 47A were reactedaccording to General Method 1 with 70.4 mg (0.204 mmol) ofmeta-chloroperbenzoic acid. Enantiomer separation of 108 mg of theracemate according to Method 9D gave 37.1 mg of the title compound fromExample 46 (Enantiomer 1) and 41.8 mg of the title compound from Example47 (Enantiomer 2).

LC-MS (Method 5B): R_(t)=0.96 min; MS (ESIpos): m/z=515 [M+H]⁺;

HPLC (Method 8E): R_(t)=5.48 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.38-7.29 (m, 4H), 4.00 (d, 1H), 3.72-3.48(m, 9H), 3.46-3.37 (m, 1H), 3.23 (s, 3H), 3.10-2.85 (m, 7H), 2.76-2.65(m, 3H), 2.32 (d, 1H), 1.98 (q, 1H).

Example 47{3-[3-(2-Methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

113 mg (0.227 mmol) of the compound from Example 47A were reactedaccording to General Method 1 with 70.4 mg (0.204 mmol) ofmeta-chloroperbenzoic acid. Enantiomer separation of 108 mg of theracemate according to Method 9D gave 37.1 mg of the title compound fromExample 46 (Enantiomer 1) and 41.8 mg of the title compound from Example47 (Enantiomer 2).

LC-MS (Method 5B): R_(t)=0.96 min; MS (ESIpos): m/z=515 [M+H]⁺;

HPLC (Method 8E): R_(t)=7.15 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): 6=7.38-7.29 (m, 4H), 4.00 (d, 1H), 3.72-3.48(m, 9H), 3.46-3.37 (m, 1H), 3.23 (s, 3H), 3.10-2.85 (m, 7H), 2.76-2.65(m, 3H), 2.32 (d, 1H), 1.98 (q, 1H).

Example 48(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

113 mg (0.227 mmol) of the compound from Example 47A were reactedaccording to General Method 2 with 196 mg (0.567 mmol) ofmeta-chloroperbenzoic acid. Enantiomer separation of 121 mg of theracemate according to Method 9D gave 34.4 mg of the title compound fromExample 48 (Enantiomer 1) and 29.2 mg of the title compound from Example49 (Enantiomer 2).

LC-MS (Method 7B): R_(t)=2.17 min; MS (ESIpos): m/z=531 [M+H]⁺;

HPLC (Method 8E): R_(t)=4.34 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.39-7.29 (m, 4H), 4.03 (d, 1H), 3.72-3.56(m, 10H), 3.46-3.36 (m, 1H), 3.23 (s, 3H), 3.18 (br. s., 4H), 3.11-2.90(m, 5H), 2.33-2.27 (m, 1H), 1.97 (q, 1H).

Example 49(1,1-Dioxidothiomorpholin-4-yl){3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

113 mg (0.227 mmol) of the compound from Example 47A were reactedaccording to General Method 2 with 196 mg (0.567 mmol) ofmeta-chloroperbenzoic acid. Enantiomer separation of 121 mg of theracemate according to Method 9D gave 34.4 mg of the title compound fromExample 48 (Enantiomer 1) and 29.2 mg of the title compound from Example49 (Enantiomer 2).

LC-MS (Method 7B): R_(t)=2.18 min; MS (ESIpos): m/z=531 [M+H]⁺;

HPLC (Method 8E): R_(t)=7.86 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.39-7.29 (m, 4H), 4.03 (d, 1H), 3.72-3.56(m, 10H), 3.46-3.36 (m, 1H), 3.23 (s, 3H), 3.18 (br. s., 4H), 3.11-2.90(m, 5H), 2.33-2.27 (m, 1H), 1.97 (q, 1H).

Example 50{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(1,1-difluoroethyl)phenyl]piperidin-1-yl}-(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

34.1 mg (0.074 mmol) of the compound from Example 54A were reactedaccording to General Method 1 with 22.9 mg (0.066 mmol) ofmeta-chloroperbenzoic acid. Yield: 39.7 mg (100% of theory).

LC-MS (Method 5B): R_(t)=0.99 min; MS (ESIpos): m/z=479 [M+H]⁺.

Example 51{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(1,1-difluoroethyl)phenyl]piperidin-1-yl}-(1-oxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 35.5 mg of the racemate from Example 50according to Method 9D gave 12.0 mg of the title compound from Example51 (Enantiomer 1) and 14.0 mg of the title compound from Example 52(Enantiomer 2).

LC-MS (Method 5B): R_(t)=1.00 min; MS (ESIpos): m/z=479 [M+H]⁺;

HPLC (Method 9E): R_(t)=5.27 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.53 (d, 2H), 7.45 (d, 2H), 3.96 (d, 1H),3.71-3.46 (m, 5H), 3.42-3.35 (m, 1H), 3.09-2.84 (m, 5H), 2.71 (d, 2H),2.29 (d, 1H), 2.16-2.08 (m, 1H), 2.02-1.90 (m, 4H), 1.10-1.02 (m, 2H),0.93-0.85 (m, 2H).

Example 52{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(1,1-difluoroethyl)phenyl]piperidin-1-yl}-(1-oxido-thiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 35.5 mg of the racemate from Example 50according to Method 9D gave 12.0 mg of the title compound from Example51 (Enantiomer 1) and 14.0 mg of the title compound from Example 52(Enantiomer 2).

LC-MS (Method 5B): R_(t)=1.00 min; MS (ESIpos): m/z=479 [M+H]⁺;

HPLC (Method 9E): R_(t)=6.78 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.53 (d, 2H), 7.45 (d, 2H), 3.96 (d, 1H),3.71-3.46 (m, 5H), 3.42-3.35 (m, 1H), 3.09-2.84 (m, 5H), 2.71 (d, 2H),2.29 (d, 1H), 2.16-2.08 (m, 1H), 2.02-1.90 (m, 4H), 1.10-1.02 (m, 2H),0.93-0.85 (m, 2H).

Example 53{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(1,1-difluoroethyl)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[racemic cis isomer]

34.1 mg (0.074 mmol) of the compound from Example 54A were reactedaccording to General Method 2 with 63.6 mg (0.184 mmol) ofmeta-chloroperbenzoic acid. Yield: 37.1 mg (99% of theory)

LC-MS (Method 5B): R_(t)=1.06 min; MS (ESIpos): m/z=495 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.52 (d, 2H), 7.44 (d, 2H), 4.00 (d, 1H),3.69-3.56 (m, 5H), 3.41-3.34 (m, 1H), 3.17 (br. s., 4H), 3.10-2.95 (m,3H), 2.28 (d, 1H), 2.17-2.07 (m, 1H), 2.03-1.89 (m, 4H), 1.10-1.01 (m,2H), 0.94-0.85 (m, 2H).

Example 54{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(1,1-difluoroethyl)phenyl]piperidin-1-yl}-(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 37.1 mg of the racemate from Example 53according to Method 9D gave 13.0 mg of the title compound from Example54 (Enantiomer 1) and 14.0 mg of the title compound from Example 55(Enantiomer 2).

HPLC (Method 9E): R_(t)=5.81 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.52 (d, 2H), 7.44 (d, 2H), 4.00 (d, 1H),3.69-3.56 (m, 5H), 3.41-3.34 (m, 1H), 3.17 (br. s., 4H), 3.10-2.95 (m,3H), 2.28 (d, 1H), 2.17-2.07 (m, 1H), 2.03-1.89 (m, 4H), 1.10-1.01 (m,2H), 0.94-0.85 (m, 2H).

Example 55{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(1,1-difluoroethyl)phenyl]piperidin-1-yl}-(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

Enantiomer separation of 37.1 mg of the racemate from Example 53according to Method 9D gave 13.0 mg of the title compound from Example54 (Enantiomer 1) and 14.0 mg of the title compound from Example 55(Enantiomer 2).

HPLC (Method 9E): R_(t)=9.63 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.52 (d, 2H), 7.44 (d, 2H), 4.00 (d, 1H),3.69-3.56 (m, 5H), 3.41-3.34 (m, 1H), 3.17 (br. s., 4H), 3.10-2.95 (m,3H), 2.28 (d, 1H), 2.17-2.07 (m, 1H), 2.03-1.89 (m, 4H), 1.10-1.01 (m,2H), 0.94-0.85 (m, 2H).

Example 56{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(difluoromethoxy)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[racemic cis isomer]

100 mg (0.23 mmol) of the compound from Example 37A and 46 mg (0.46mmol) of N-hydroxycyclopropanecarboximidamide were initially charged in0.8 ml of DMF and reacted with 132 mg (0.35 mmol) of HATU and 0.12 ml(90 mg, 0.69 mmol) of N,N-diisopropylethylamine. The reaction mixturewas stirred at RT for 15 minutes and then partitioned between water andethyl acetate. The organic phase was washed repeatedly with water, driedover sodium sulphate and concentrated under reduced pressure. Theresidue was taken up in 3.0 ml of DMF and converted in a microwave at180° C. for 2 minutes. The reaction mixture was purified by means ofpreparative HPLC. Yield: 46 mg (37% of theory)

LC-MS (Method 2B): R_(t)=1.20 min; MS (ESIpos): m/z=497 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.43-7.30 (m, 2H), 7.14 (d, 2H), 4.09 (q,1H), 3.99 (br. d, 1H), 3.63 (br. d., 1H), 3.40-3.33 (m, 1H), 3.33-3.28(m, 4H), 3.22-3.10 (m, 4H), 3.08-2.88 (m, 3H), 2.28 (br. d, 1H),2.16-2.07 (m, 1H), 2.00-1.87 (m, 1H), 1.12-0.99 (m, 2H), 0.94-0.84 (m,2H).

Example 57{3-[4-(Difluoromethoxy)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[racemic cis isomer]

300 mg (0.69 mmol) of the compound from Example 37A and 246 mg (2.08mmol) of N′-hydroxy-3-methoxypropanimidamide were initially charged in2.6 ml of DMF and reacted with 396 mg (1.0 mmol) of HATU and 0.36 ml(269 mg, 2.1 mmol) of N,N-diisopropylethylamine. The reaction mixturewas stirred at RT for 15 minutes and then partitioned between water andethyl acetate. The organic phase was washed repeatedly with water, driedover sodium sulphate and concentrated under reduced pressure. Theresidue was dissolved in 2.0 ml of DMF and converted in a microwave at180° C. for 2 minutes. The reaction mixture was purified by means ofpreparative

HPLC. Yield: 141 mg (38% of theory)

LC-MS (Method 2B): R_(t)=1.10 min; MS (ESIpos): m/z=515 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): 6=7.43-7.33 (m, 2H), 7.15 (d, 2H), 4.11-3.99(m, 2H), 3.71-3.64 (m, 3H), 3.64-3.55 (m, 4H), 3.46-3.35 (m, 1H),3.35-3.30 (m, 4H), 3.18 (br. s, 3H), 3.14-2.90 (m, 5H), 2.30 (br. d,1H), 2.03-1.92 (m, 1H).

Example 58{3-[4-(Difluoromethoxy)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

The enantiomer separation of 117 mg of the racemate from Example 57according to Method 10D gave 43 mg of the compound from Example 58(Enantiomer 1) and 38 mg of the compound from Example 59 (Enantiomer 2).

HPLC (Method 10E): R_(t)=4.17 min, >99.0% ee;

LC-MS (Method 2B): R_(t)=1.10 min; MS (ESIpos): m/z=515 [M+H]⁺.

Example 59{3-[4-(Difluoromethoxy)phenyl]-5-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[enantiomerically pure cis isomer]

The enantiomer separation of 117 mg of the racemate from Example 57according to Method 10D gave 43 mg of the compound from Example 58(Enantiomer 1) and 38 mg of the compound from Example 59 (Enantiomer 2).

HPLC (Method 10E): R_(t)=9.24 min, >99.0% ee;

LC-MS (Method 2B): R_(t)=1.10 min; MS (ESIpos): m/z=515 [M+H]⁺.

Example 60{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(difluoromethoxy)phenyl]piperidin-1-yl}(1-oxidothiomorpholin-4-yl)methanone[racemic cis isomer]

200 mg (0.48 mmol) of the compound from Example 39A and 96 mg (0.96mmol) of N′-hydroxycyclopropanecarboximidamide were initially charged in1.8 ml of DMF and reacted with 274 mg (0.72 mmol) of HATU and 0.25 ml(186 mg, 1.44 mmol) of N,N-diisopropylethylamine. The reaction mixturewas stirred at RT for 15 minutes and then partitioned between water andethyl acetate. The organic phase was washed repeatedly with water, driedover sodium sulphate and concentrated under reduced pressure. Theresidue was dissolved in 2.0 ml of DMF and converted in a microwave at180° C. for 2 minutes. The reaction mixture was purified by means ofpreparative HPLC. Yield: 25 mg (10% of theory)

LC-MS (Method 2B): R_(t)=1.12 min; MS (ESIpos): m/z=481 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.42-7.35 (m, 2H), 7.14 (d, 2H), 4.01-3.87(m, 1H), 3.69-3.45 (m, 5H), 3.42-3.34 (m, 1H), 3.07-2.85 (m, 5H), 2.70(br. d, 2H), 2.34-2.23 (m, 1H), 2.15-2.07 (m, 1H), 1.99-1.88 (m, 1H),1.12-1.01 (m, 2H), 0.94-0.85 (m, 2H).

Example 61(1,1-Dioxidothiomorpholin-4-yl){3-[3-(1-methoxycyclopropyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}methanone[racemic cis isomer]

29.0 mg (0.162 mmol) of the compound from Example 65A were reactedaccording to General Method 2 with 49.0 mg (0.142 mmol) ofmeta-chloroperbenzoic acid. Yield: 31.2 mg (95% of theory)

LC-MS (Method 5B): R_(t)=1.06 min; MS (ESIpos): m/z=543 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ=7.37-7.29 (m, 4H), 4.02 (d, 1H), 3.68-3.55(q, 7H), 3.38 (s, 3H), 3.17 (br. s., 4H), 3.10-2.88 (m, 3H), 2.30 (d,1H), 1.95 (q, 1H), 1.34-1.28 (m, 2H), 1.20-1.12 (m, 2H).

Example 62(1,1-Dioxidothiomorpholin-4-yl){3-[3-(1-methoxycyclopropyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

Enantiomer separation of 31.2 mg of the racemate from Example 61according to Method 11D gave 12.0 mg of the title compound from Example62 (Enantiomer 1) and 12.0 mg of the title compound from Example 63(Enantiomer 2).

LC-MS (Method 2B): R_(t)=1.26 min; MS (ESIpos): m/z=543 [M+H]⁺;

HPLC (Method 11E): R_(t)=17.9 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.37-7.29 (m, 4H), 4.02 (d, 1H), 3.68-3.55(q, 7H), 3.38 (s, 3H), 3.17 (br. s., 4H), 3.10-2.88 (m, 3H), 2.30 (d,1H), 1.95 (q, 1H), 1.34-1.28 (m, 2H), 1.20-1.12 (m, 2H).

Example 63(1,1-Dioxidothiomorpholin-4-yl){3-[3-(1-methoxycyclopropyl)-1,2,4-oxadiazol-5-yl]-5-[4-(2,2,2-trifluoroethyl)phenyl]piperidin-1-yl}methanone[enantiomerically pure cis isomer]

Enantiomer separation of 31.2 mg of the racemate from Example 61according to Method 11D gave 12.0 mg of the title compound from Example62 (Enantiomer 1) and 12.0 mg of the title compound from Example 63(Enantiomer 2).

LC-MS (Method 2B): R_(t)=1.26 min; MS (ESIpos): m/z=543 [M+H]⁺;

HPLC (Method 11E): R_(t)=29.2 min, >99.0% ee;

¹H NMR (400 MHz, DMSO-d₆): δ=7.37-7.29 (m, 4H), 4.02 (d, 1H), 3.68-3.55(q, 7H), 3.38 (s, 3H), 3.17 (br. s., 4H), 3.10-2.88 (m, 3H), 2.30 (d,1H), 1.95 (q, 1H), 1.34-1.28 (m, 2H), 1.20-1.12 (m, 2H).

B) ASSESSMENT OF PHYSIOLOGICAL ACTIVITY Abbreviations:

BSA bovine serum albumin

DMEM Dulbecco's Modified Eagle Medium

EGTA ethylene glycol bis-(2-aminoethyl)-N,N,N′,N′-tetraacetic acidFCS fetal calf serumHEPES 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid[3H]haTRAP tritiated high affinity thrombin receptor activating peptidePRP platelet-rich plasma

The suitability of the inventive compounds for treating thromboembolicdisorders can be demonstrated in the following assay systems:

1.) In Vitro Assays 1.a) Cellular Functional In Vitro Test

A recombinant cell line is used to identify agonists of the humanprotease activated receptor 1 (PAR1) and to quantify the activity of thesubstances described herein. The cell is originally derived from a humanembryonal kidney cell (HEK293; ATCC: American Type Culture Collection,Manassas, Va. 20108, USA). The test cell line constitutively expresses amodified form of the calcium-sensitive photoprotein aequorin which,after reconstitution with the cofactor coelenterazine, emits light whenthe free calcium concentration in the inner mitochondrial compartment isincreased (Rizzuto R, Simpson A W, Brini M, Pozzan T.; Nature 1992, 358,325-327). Additionally, the cell stably expresses the endogenous humanPAR1 receptor and the endogenous purinergic receptor P2Y2. The resultingPAR1 test cell responds to stimulation of the endogenous PAR1 or P2Y2receptor with an intracellular release of calcium ions, which can bequantified through the resulting aequorin luminescence with a suitableluminometer (Milligan G, Marshall F, Rees S, Trends in PharmacologicalSciences 1996, 17, 235-237).

For the testing of the substance specificity, the effect thereof afteractivation of the endogenous PAR1 receptor is compared with the effectafter activation of the endogenous purinergic P2Y2 receptor whichutilizes the same intracellular signal path.

Test Procedure:

The cells are plated out two days (48 hours) before the test in culturemedium (DMEM F12, supplemented with 10% FCS, 2 mM glutamine, 20 mMHEPES, 1.4 mM pyruvate, 0.1 mg/ml gentamycin, 0.15% sodium bicarbonate;BioWhittaker Cat. #BE04-687Q; B-4800 Verviers, Belgium) in 384-wellmicrotitre plates and kept in a cell incubator (96% atmospherichumidity, 5% v/v CO₂, 37° C.). On the day of the test, the culturemedium is replaced by a Tyrode's solution (in mM: 140 sodium chloride, 5potassium chloride, 1 magnesium chloride, 2 calcium chloride, 20glucose, 20 HEPES), which additionally contains the cofactorcoelenterazine (25 μM) and glutathione (4 mM), and the microtitre plateis then incubated for a further 3-4 hours. The test substances are thenpipetted onto the microtitre plate, and 5 minutes after the transfer ofthe test substances into the wells of the microtitre plate the plate istransferred into the luminometer, a PAR1 agonist concentration whichcorresponds to EC₅₀ is added and the resulting light signal isimmediately measured in the luminometer. To distinguish an antagonistsubstance action from a toxic action, the endogenous purinergic receptoris immediately subsequently activated with agonist (ATP, finalconcentration 10 μM) and the resulting light signal is measured. Theresults are shown in Table A:

TABLE A Example No. IC₅₀ [nM] 1 43 8 33 10 8.0 15 5.1 20 23 31 32 52 4.754 4.3 61 15.7

1.b) PAR-1 Receptor Binding Assay

Platelet membranes are incubated with 12 nM [3H]haTRAP and testsubstance in different concentrations in a buffer (50 mM Tris pH 7.5, 10mM magnesium chloride, 1 mM EGTA, 0.1% BSA) at room temperature for 80mM Then the mixture is transferred to a filter plate and washed twicewith buffer. After addition of scintillation liquid, the radioactivityon the filter is measured in a beta counter.

1.c) Platelet Aggregation in Plasma

Platelet aggregation is determined using blood from healthy volunteersof both genders, who had not received any thrombocyteaggregation-influencing medication for the last ten days. The blood istaken up into monovettes (Sarstedt, Nümbrecht, Germany) which contain,as anticoagulant, sodium citrate 3.8% (1 part citrate+9 parts blood). Toobtain platelet-rich plasma, the citrated whole blood is centrifuged at140 g for 20 min.

For the aggregation measurements, aliquots of the platelet-rich plasmawith increasing concentrations of test substance are incubated at 37° C.for 10 min. Subsequently, aggregation is triggered by addition of athrombin receptor agonist (TRAP6, SFLLRN) in an aggregometer anddetermined at 37° C. by means of the turbidimetry method according toBorn (Born, G. V. R., Cross M. J., The Aggregation of Blood Platelets;J. Physiol. 1963, 168, 178-195). The SFLLRN concentration leading tomaximum aggregation is, if appropriate, determined individually for eachdonor.

To calculate the inhibitory effect, the maximum increase in lighttransmission (amplitude of the aggregation curve in %) is determinedwithin 5 minutes after addition of the agonist in the presence andabsence of test substance, and the inhibition is calculated. Theinhibition curves are used to calculate the concentration which inhibitsaggregation by 50%. The results are shown in Table B:

TABLE B Example No. IC₅₀ [nM]  8 0.29 10 0.49 13 0.17 52 0.58

1.d) Platelet Aggregation in Buffer

Platelet aggregation is determined using blood from healthy volunteersof both genders, who had not received any thrombocyteaggregation-influencing medication for the last ten days. The blood istaken up into monovettes (Sarstedt, Nümbrecht, Germany) which contain,as anticoagulant, sodium citrate 3.8% (1 part citrate+9 parts blood). Toobtain platelet-rich plasma, the citrated whole blood is centrifuged at140 g for 20 mM One quarter of the volume of ACD buffer (44.8 mM sodiumcitrate, 20.9 mM citric acid, 74.1 mM glucose and 4 mM potassiumchloride) is added to the PRP, and the mixture is centrifuged at 1000 gfor 10 minutes. The platelet pellet is resuspended in wash buffer andcentrifuged at 1000 g for 10 minutes. The platelets are resuspended inincubation buffer and adjusted to 200 000 cells/W. Prior to the start ofthe test, calcium chloride and magnesium chloride, final concentrationin each case 2 mM (2M stock solution, dilution 1:1000), are added. Note:in the case of ADP-induced aggregation, only calcium chloride is added.The following agonists can be used: TRAP6-trifluoroacetate salt,collagen, human α-thrombin and U-46619. For each donor, theconcentration of the agonist is tested.

Test Procedure:

96-well microtitre plates are used. The test substance is diluted inDMSO, and 2 μl per well is initially charged. 178 μl of plateletsuspension are added, and the mixture is preincubated at roomtemperature for 10 minutes. 20 μl of agonist are added, and themeasurement in the Spectramax, OD 405 nm, is started immediately.Kinetics are determined in 11 measurements of 1 minute each. Between themeasurements, the mixture is shaken for 55 seconds.

1.e) Platelet Aggregation in Fibrinogen-Depleted Plasma

To determine platelet aggregation, blood of healthy volunteers of bothgenders, who had not received any thrombocyte aggregation-influencingmedication for the last ten days, is used. The blood is taken up intomonovettes (Sarstedt, Nümbrecht, Germany) which contain, asanticoagulant, sodium citrate 3.8% (1 part citrate+9 parts blood).

1.e) Preparation of Fibrinogen-Depleted Plasma:

To obtain low-platelet plasma, the citrated whole blood is centrifugedat 140 g for 20 min. The low-platelet plasma is admixed in a ratio of1:25 with reptilase (Roche Diagnostic, Germany) and inverted cautiously.This is followed by 10 mM of incubation at 37° C. in a water bath,followed directly by 10 mM of incubation on ice. The plasma/reptilasemixture is centrifuged at 1300 g for 15 mM, and the supernatant(fibrinogen-depleted plasma) is obtained.

Platelet Isolation:

To obtain platelet-rich plasma, the citrated whole blood is centrifugedat 140 g for 20 min. One quarter of the volume of ACD buffer (44.8 mMsodium citrate, 20.9 mM citric acid, 74.1 mM glucose and 4 mM potassiumchloride) is added to the PRP, and the mixture is centrifuged at 1300 gfor 10 minutes. The platelet pellet is resuspended in wash buffer andcentrifuged at 1300 g for 10 minutes. The platelets are resuspended inincubation buffer and adjusted to 400 000 cells/4 and calcium chloridesolution is added to a final concentration of 5 mM (dilution 1/200).

For the aggregation measurements, aliquots (98 μl of fibrinogen-depletedplasma and 80 μl of platelet suspension) are incubated with increasingconcentrations of test substance at RT for 10 mM Subsequently,aggregation is triggered by addition of human alpha thrombin in anaggregometer and determined at 37° C. by means of the turbidimetrymethod according to Born (Born, G. V. R., Cross M. J., The Aggregationof Blood Platelets; J. Physiol. 1963, 168, 178-195). The alpha thrombinconcentration which just leads to the maximum aggregation is determinedindividually for each donor.

To calculate the inhibitory activity, the increase in the maximum lighttransmission (amplitude of the aggregation curve in %) is determinedwithin 5 minutes after addition of the agonist in the presence andabsence of test substance, and the inhibition is calculated. Theinhibition curves are used to calculate the concentration which inhibitsaggregation by 50%.

1.f) Stimulation of Washed Platelets and Analysis in Flow Cytometry

Isolation of Washed Platelets:

Human whole blood is obtained by venipuncture from voluntary donors andtransferred into monovettes (Sarstedt, Nümbrecht, Germany) containingsodium citrate as anticoagulant (1 part sodium citrate 3.8%+9 partswhole blood). The monovettes are centrifuged at 90° rotations per minuteand 4° C. for a period of 20 minutes (Heraeus Instruments, Germany;Megafuge LORS). The platelet-rich plasma is cautiously removed andtransferred into a 50 ml Falcon tube. ACD buffer (44 mM sodium citrate,20.9 mM citric acid, 74.1 mM glucose) is then added to the plasma. Thevolume of the ACD buffer corresponds to one quarter of the plasmavolume. Centrifuging at 2500 rpm and 4° C. for ten minutes sediments theplatelets. Thereafter, the supernatant is cautiously decanted off anddiscarded. The precipitated platelets are first cautiously resuspendedin one millilitre of wash buffer (113 mM sodium chloride, 4 mM disodiumhydrogenphosphate, 24 mM sodium dihydrogenphosphate, 4 mM potassiumchloride, 0.2 mM ethylene glycol bis(2-aminoethyl)-N,N,N′,N′-tetraaceticacid, 0.1% glucose) and then made up with wash buffer to a volume whichcorresponds to that of the amount of plasma. The wash procedure isrepeated. The platelets are precipitated by another ten-minutecentrifugation at 2500 rpm and 4° C. and then carefully resuspended inone millilitre of incubation buffer (134 mM sodium chloride, 12 mMsodium hydrogencarbonate, 2.9 mM potassium chloride, 0.34 mM sodiumdihydrogencarbonate, 5 mM HEPES, 5 mM glucose, 2 mM calcium chloride and2 mM magnesium chloride) and adjusted with incubation buffer to aconcentration of 300 000 platelets per μl.

Staining and Stimulation of the Human Platelets with Human α-Thrombin inthe Presence or Absence of a PAR-1 Antagonist:

The platelet suspension is preincubated with the substance to be testedor the appropriate solvent at 37° C. for 10 minutes (Eppendorf, Germany;Thermomixer Comfort). Platelet activation is triggered by addition ofthe agonist (0.5 μM or 1 μM α-thrombin; Kordia, the Netherlands, 3281NIH units/mg; or 30 μg/ml of thrombin receptor activating peptide(TRAP6); Bachem, Switzerland) at 37° and with shaking at 500 rpm. Ateach of 0, 1, 2.5, 5, 10 and 15 minutes, one aliquot of 50 μl is removedand transferred into one millilitre of singly concentrated CellFix™solution (Becton Dickinson Immunocytometry Systems, USA). To fix thecells, they are incubated in the dark at 4° C. for 30 minutes. Theplatelets are precipitated by centrifuging at 600 g and 4° C. for tenminutes. The supernatant is discarded and the platelets are resuspendedin 400 μl CellWash™ (Becton Dickinson Immunocytometry Systems, USA). Onealiquot of 100 μl is transferred to a new FACS tube. 1 μl of theplatelet-identifying antibody and 1 μl of the activation state-detectingantibody are made up to a volume of 100 μl with CellWash™. This antibodysolution is then added to the platelet suspension and incubated in thedark at 4° C. for 20 minutes. After staining, the mixture volume isincreased by addition of a further 400 μl of CellWash™

The platelets are identified using a fluoresceinisothiocyanate-conjugated antibody directed against human glycoproteinIIb (CD41) (Immunotech Coulter, France; Cat. No. 0649). With the aid ofthe phycoerythrin-conjugated antibody directed against humanglycoprotein P-selectin (Immunotech Coulter, France; Cat. No. 1759), itis possible to determine the activation state of the platelets.P-selectin (CD62P) is localized in the α-granules of resting platelets.However, following in vitro or in vivo stimulation, it is translocalizedto the external plasma membrane.

Flow Cytometry and Data Evaluation:

The samples are analysed in the FACSCalibur™ Flow Cytometry Systeminstrument from Becton Dickinson Immunocytometry Systems, USA, andevaluated and graphically represented with the aid of the CellQuestsoftware, Version 3.3 (Becton Dickinson Immunocytometry Systems, USA).The degree of platelet activation is determined by the percentage ofCD62P-positive platelets (CD41-positive events). From each sample, 10000 CD41-positive events are counted.

The inhibitory effect of the substances to be tested is calculated viathe reduction in platelet activation, which relates to the activation bythe agonist.

1.g) Platelet Aggregation Measurement Using the Parallel-Plate FlowChamber

Platelet aggregation is determined using blood from healthy volunteersof both genders, who had not received any thrombocyteaggregation-influencing medication for the last ten days. The blood istaken up into monovettes (Sarstedt, Nümbrecht, Germany) which contain,as anticoagulant, sodium citrate 3.8% (1 part citrate+9 parts blood). Toobtain platelet-rich plasma, the citrated whole blood is centrifuged at140 g for 20 mM One quarter of the volume of ACD buffer (44.8 mM sodiumcitrate, 20.9 mM citric acid, 74.1 mM glucose and 4 mM potassiumchloride) is added to the PRP, and the mixture is centrifuged at 1000 gfor 10 minutes. The platelet pellet is resuspended in wash buffer andcentrifuged at 1000 g for 10 minutes. For the perfusion study, a mixtureof 40% erythrocytes and 60% washed platelets (200 000/μl) is preparedand suspended in HEPES-tyrode buffer. Platelet aggregation under flowconditions is measured using the parallel-plate flow chamber (B.Nieswandt et al., EMBO J. 2001, 20, 2120-2130; C. Weeterings,Arterioscler Thromb. Vasc. Biol. 2006, 26, 670-675; J J Sixma, Thromb.Res. 1998, 92, 43-46). Glass slides are wetted with 100 μl of a solutionof human α-thrombin (dissolved in Tris buffer) at 4° C. overnight(α-thrombin in different concentrations, for example 10 to 50 μg/ml) andfinally blocked using 2% BSA.

Reconstituted blood is passed over the thrombin-wetted glass slides at aconstant flow rate (for example a shear rate of 300/second) for 5minutes and observed and recorded using a microscope video system. Theinhibitory effect of the substances to be tested is determinedmorphometrically via the reduction in platelet aggregate formation.Alternatively, the inhibition of the platelet activation can bedetermined by flow cytometry, for example via p-selectin expression(CD62p) (see Method 1.f).

1.h) Platelet Aggregation and Activation Measurement Using theParallel-Plate Flow Chamber (Anticoagulated Blood, Collagen)

Platelet activation under flow conditions is determined using blood fromhealthy volunteers of both genders, who had not received any thrombocyteaggregation-influencing medication for the last ten days. The blood istaken up into monovettes (Sarstedt, Númbrecht, Germany) which contain,as anticoagulant, sodium citrate 3.8% (1 part citrate+9 parts blood).

The measurement of platelet activation is carried out using theparallel-plate flow chamber (B. Nieswandt et al., EMBO J. 2001, 20,2120-2130; C. Weeterings, Arterioscler Thromb. Vasc. Biol. 2006, 26,670-675; J J Sixma, Thromb. Res. 1998, 92, 43-46). Glass slides arewetted with 20 μl of collagen suspension (collagen reagent: Horm,Nycomed) at 4° C. overnight (type I collagen in differentconcentrations, e.g. 1-10 ng/slide) and finally blocked using 2% BSA.

To prevent fibrin clot formation, citrated whole blood is admixed withPefabloc FG (Pentapharm, final concentration 3 mM) and, by addition ofCaCl₂ solution (final Ca⁺⁺ concentration 5 mM), passed over thecollagen-coated glass slides at a constant flow rate (for example ashear rate of 1000/second) for 5 minutes and observed and recorded usinga microscope video system. The inhibitory effect of the substances to betested is determined morphometrically via the reduction in plateletaggregate formation. Alternatively, the inhibition of the plateletactivation can be determined by flow cytometry, for example viap-selectin expression (CD62p) (see Method 1.0.

1.i) Platelet Aggregation and Activation Measurement Using theParallel-Plate Flow Chamber (Nonanticoagulated Blood, Collagen)

Platelet activation under flow conditions is determined using blood fromhealthy volunteers of both genders, who had not received any thrombocyteaggregation-influencing medication for the last ten days. The blood istaken up into neutral monovettes (Sarstedt, Nümbrecht, Germany) which donot contain any anticoagulant, and immediately admixed with Pefabloc FG(Pentapharm, final concentration 3 mM) to prevent fibrin clot formation.Test substances dissolved in DMSO are added simultaneously withPefablock FG and introduced without further incubation into theparallel-plate flow chamber. The measurement of platelet activation isconducted by morphometry or flow cytometry in the collagen-coatedparallel-plate flow chamber, as described in Method 1.h).

2.) Ex Vivo Assay 2.a) Platelet Aggregation (Primates, Guinea Pigs)

Awake or anaesthetized guinea pigs or primates are treated orally,intravenously or intraperitoneally with test substances in suitableformulations. As a control, other guinea pigs or primates are treated inan identical manner with the corresponding vehicle. Depending on themode of application, blood is obtained from the deeply anaesthetizedanimals by puncture of the heart or of the aorta for different times.The blood is transferred into monovettes (Sarstedt, Nümbrecht, Germany)which, as anticoagulant, contain sodium citrate 3.8% (1 part citratesolution+9 parts blood). To obtain platelet-rich plasma, the citratedwhole blood is centrifuged at 140 g for 20 min.

Aggregation is triggered by addition of a thrombin receptor agonist(TRAP6, SFLLRN, 50 μg/ml; in each experiment, the concentration isdetermined for each animal species) in an aggregometer and determinedusing the turbidimetry method according to Born (Born, G. V. R., CrossM. J., The Aggregation of Blood Platelets; J. Physiol. 1963, 168,178-195) at 37° C.

To measure the aggregation, the maximum increase in the lighttransmission (amplitude of the aggregation curve in %) is determined 5minutes after addition of the agonist. The inhibitory effect of theadministered test substances in the treated animals is calculated viathe reduction in aggregation, based on the mean of the control animals.

In addition to measurement of aggregation, the inhibition of plateletactivation can be determined by flow cytometry, for example viap-selectin expression (CD62p) (see Method 1.f).

2.b) Platelet Aggregation and Activation Measurement in theParallel-Plate Flow Chamber (Primates)

Awake or anaesthetized primates are treated orally, intravenously orintraperitoneally with test substances in suitable formulations. As acontrol, other animals are treated in an identical manner with thecorresponding vehicle. According to the mode of administration, blood isobtained from the animals by venipuncture for different times. The bloodis transferred into monovettes (Sarstedt, Nümbrecht, Germany) which, asanticoagulant, contain sodium citrate 3.8% (1 part citrate solution+9parts blood). Alternatively, nonanticoagulated blood can be taken withneutral monovettes (Sarstedt). In both cases, the blood is admixed withPefabloc FG (Pentapharm, final concentration 3 mM) to prevent fibrinclot formation.

Citrated whole blood is recalcified before the measurement by addingCaCl₂ solution (final Ca⁺⁺ concentration 5 mM). Nonanticoagulated bloodis introduced directly into the parallel-plate flow chamber foranalysis. The measurement of platelet activation is conducted bymorphometry or flow cytometry in the collagen-coated parallel-plate flowchamber, as described in Method 1.h).

3.) In Vivo Assays 3.a) Thrombosis Model

The inventive compounds can be studied in thrombosis models in suitableanimal species in which thrombin-induced platelet aggregation ismediated via the PAR-1 receptor. Suitable animal species are guinea pigsand, in particular, primates (cf.: Lindahl, A. K., Scarborough, R. M.,Naughton, M. A., Harker, L. A., Hanson, S. R., Thromb Haemost 1993, 69,1196; Cook J J, Sitko G R, Bednar B, Condra C, Mellott M J, Feng D-M,Nutt R F, Shager J A, Gould R J, Connolly T M, Circulation 1995, 91,2961-2971; Kogushi M, Kobayashi H, Matsuoka T, Suzuki S, Kawahara T,Kajiwara A, Hishinuma I, Circulation 2003, 108 Suppl. 17, IV-280; DerianC K, Damiano B P, Addo M F, Darrow A L, D'Andrea M R, Nedelman M, ZhangH-C, Maryanoff B E, Andrade-Gordon P, J. Pharmacol. Exp. Ther. 2003,304, 855-861). Alternatively, it is possible to use guinea pigs whichhave been pretreated with inhibitors of PAR-3 and/or PAR-4 (Leger A J etal., Circulation 2006, 113, 1244-1254), or transgenic PAR-3- and/orPAR-4-knockdown guinea pigs.

3.b) Impaired Coagulation and Organ Dysfunction in the Case ofDisseminated Intravasal Coagulation (DIC)

The inventive compounds can be studied in models of DIC and/or sepsis insuitable animal species. Suitable animal species are guinea pigs and, inparticular, primates, and for the study of endothelium-mediated effectsalso mice and rats (cf.: Kogushi M, Kobayashi H, Matsuoka T, Suzuki S,Kawahara T, Kajiwara A, Hishinuma I, Circulation 2003, 108 Suppl. 17,IV-280; Derian C K, Damiano B P, Addo M F, Darrow A L, D'Andrea M R,Nedelman M, Zhang H-C, Maryanoff B E, Andrade-Gordon P, J. Pharmacol.Exp. Ther. 2003, 304, 855-861; Kaneider N C et al., Nat Immunol, 2007,8, 1303-12; Camerer E et al., Blood, 2006, 107, 3912-21; Riewald M etal., J Biol Chem, 2005, 280, 19808-14.). Alternatively, it is possibleto use guinea pigs which have been pretreated with inhibitors of PAR-3and/or PAR-4 (Leger A J et al., Circulation 2006, 113, 1244-1254), ortransgenic PAR-3- and/or PAR-4-knockdown guinea pigs.

3.b.1) Thrombin-Antithrombin Complexes

Thrombin-antithrombin complexes (referred to hereinafter as “TAT”) are ameasure of the thrombin formed endogenously by coagulation activation.TATs are determined via an ELISA assay (Enzygnost TAT micro,Dade-Behring). Plasma is obtained from citrated blood by centrifugation.50 μl of TAT sample buffer are added to 50 μl of plasma, shaken brieflyand incubated at room temperature for 15 min The samples are filteredwith suction, and the well is washed 3 times with wash buffer (300μl/well). Between the wash steps, the plate is tapped to remove anyresidual wash buffer. Conjugate solution (100 μl) is added and themixture is incubated at room temperature for 15 min The samples arefiltered with suction, and the well is washed 3 times with wash buffer(300 μl/well). The chromogenic substrate (100 μl/well) is then added,the mixture is incubated in the dark at room temperature for 30 min,stop solution (100 μl/well) is added, and the development of colour at492 nm is measured (Safire plate reader).

3.b.2) Parameters of Organ Dysfunction

Various parameters are determined, which allow conclusions to be drawnwith respect to the functional restriction of various internal organsowing to the administration of LPS, and the therapeutic effect of testsubstances can be assessed. Citrated blood or, if appropriate, lithiumheparin blood, is centrifuged, and the plasma is used to determine theparameters. Typically, the following parameters are determined:creatinine, urea, aspartate aminotransferase (AST), alanineaminotransferase (ALT), total bilirubin, lactate dehydrogenase (LDH),total protein, total albumin and fibrinogen. The values give informationregarding kidney function, liver function, cardiovascular function andvascular function.

3.b.3) Parameters of Inflammation

The extent of the inflammatory reaction triggered by endotoxin can bedemonstrated by the increase in inflammation mediators, for exampleinterleukins (1, 6, 8 and 10), tumour necrosis factor alpha or monocytechemoattractant protein-1, in the plasma. ELISAs or the Luminex systemmay be used for this purpose.

3.c) Antitumour Activity

The inventive compounds can be tested in models of cancer, for examplein the human breast cancer model in immunodeficient mice (cf.: S.Even-Ram et. al., Nature Medicine, 1988, 4, 909-914).

3.d) Antiangiogenetic Activity

The inventive compounds can be tested in in vitro and in vivo models ofangiogenesis (cf.: Caunt et al., Journal of Thrombosis and Haemostasis,2003, 10, 2097-2102; Haralabopoulos et al., Am J Physiol, 1997,C239-C245; Tsopanoglou et al., JBC, 1999, 274, 23969-23976; Zania etal., JPET, 2006, 318, 246-254).

3.e) Blood Pressure- and Pulse-Modulating Activity

The inventive compounds can be tested in in vivo models for their actionon arterial blood pressure and heart rate. To this end, rats (forexample Wistar) are provided with implantable radiotelemetry units, andan electronic data acquisition and storage system (Data Sciences, MN,USA) consisting of a chronically implantable transducer/transmitter unitin combination with a liquid-filled catheter is employed. Thetransmitter is implanted into the peritoneal cavity, and the sensorcatheter is positioned in the descending aorta. The inventive compoundscan be administered (for example orally or intravenously). Prior to thetreatment, the mean arterial blood pressure and the heart rate of theuntreated and treated animals are measured, and it is ensured that theyare in the range of about 131-142 mmHg and 279-321 beats/minute.PAR-1-activating peptide (SFLLRN; for example doses between 0.1 and 5mg/kg) is administered intravenously. Blood pressure and heart rate aremeasured at different time intervals and periods with and withoutPAR-1-activating peptide and with and without an inventive compound(cf.: Cicala C et al., The FASEB Journal, 2001, 15, 1433-5; Stasch J Pet al., British Journal of Pharmacology 2002, 135, 344-355).

3.f) Thrombosis Model

A further in vivo thrombosis assay which is suitable for determining theefficacy of the compounds of the present invention is described inTucker E I, Marzec U M, White T C, Hurst S, Rugonyi S, McCarty O J T,Gailani D, Gruber A, Hanson SR: Prevention of vascular graft occlusionand thrombus-associated thrombin generation by inhibition of factor XI.Blood 2009, 113, 936-944.

4.) Determination of the Solubility Preparation of the Starting Solution(Original Solution)

At least 1.5 mg of the test substance are weighed out accurately into awide-mouth 10 mm screw V-vial (from Glastechnik Grafenroda GmbH, Art.No. 8004-WM-H/V15μ) with fitting screw cap and septum, DMSO is added toa concentration of 50 mg/ml and the vial is vortexed for 30 minutes.

Preparation of the Calibration Solutions:

The pipetting steps necessary are effected in 1.2 ml 96-deep well plates(DWP) with the aid of a liquid-handling robot. The solvent used is amixture of acetonitrile/water 8:2.

Preparation of the Starting Solution of Calibration Solutions (StockSolution):

833 μl of the solvent mixture are added to 10 μl of the originalsolution (concentration=600 μg/ml), and the mixture is homogenized.1:100 dilutions in separate DWPs are prepared from each test substance,and these are homogenized in turn.

Calibration Solution 5 (600 ng/ml):

270 μl of the solvent mixture are added to 30 μl of the stock solution,and the mixture is homogenized.

Calibration Solution 4 (60 ng/ml):

270 μl of the solvent mixture are added to 30 μl of the calibrationsolution 5, and the mixture is homogenized.

Calibration Solution 3 (12 ng/ml):

400 μl of the solvent mixture are added to 100 μl of the calibrationsolution 4, and the mixture is homogenized.

Calibration Solution 2 (12 ng/ml):

270 μl of the solvent mixture are added to 30 μl of the calibrationsolution 3, and the mixture is homogenized.

Calibration Solution 1 (0.6 ng/ml):

150 μl of the solvent mixture are added to 150 μl of the calibrationsolution 2, and the mixture is homogenized.

Preparation of the Sample Solutions:

The pipetting steps necessary are effected in 1.2 ml 96-well DWPs withthe aid of a liquid-handling robot. 1000 μl of PBS buffer pH 6.5 areadded to 10.1 μl of the stock solution. (PBS buffer pH 6.5: 61.86 g ofsodium chloride, 39.54 g of sodium dihydrogenphosphate and 83.35 g of 1N aqueous sodium hydroxide solution are weighed out into a 1 litrestandard flask and made up to the mark with water, and the mixture isstirred for about 1 hour. 500 ml of this solution are introduced into a5 litre measuring flask and made up to the mark with water. The pH isadjusted to 6.5 using 1 N aqueous sodium hydroxide solution.)

Procedure:

The pipetting steps necessary are effected in 1.2 ml 96-well DWPs withthe aid of a liquid-handling robot. The sample solutions prepared inthis manner are shaken at 1400 rpm and at 20° C. using a variabletemperature shaker for 24 hours. 180 μl are removed from each of thesesolutions and transferred into Beckman Polyallomer centrifuge tubes.These solutions are centrifuged at about 223 000×g for 1 hour. From eachsample solution, 100 μl of the supernatant are removed and diluted 1:10and 1:1000 with PBS buffer 6.5.

Analysis:

The samples are analysed by means of HPLC/MS-MS. The test compound isquantified by means of a five-point calibration curve. The solubility isexpressed in mg/l. Analysis sequence: 1) blank (solvent mixture); 2)calibration solution 0.6 ng/ml; 3) calibration solution 1.2 ng/ml; 4)calibration solution 12 ng/ml; 5) calibration solution 60 ng/ml; 6)calibration solution 600 ng/ml; 7) blank (solvent mixture); 8) samplesolution 1:1000; 9) sample solution 1:10.

HPLC/MS-MS Method:

HPLC: Agilent 1100, quat. pump (G1311A), autosampler CTC HTS PAL,degasser (G1322A) and column thermostat (G1316A); column: Oasis HLB 20mm×2.1 mm, 25μ; temperature: 40° C.; eluent A: water+0.5 ml of formicacid/1; eluent B: acetonitrile+0.5 ml of formic acid/1; flow rate: 2.5ml/min; stop time 1.5 min; gradient: 0 min 95% A, 5% B; ramp: 0-0.5 min5% A, 95% B; 0.5-0.84 min 5% A, 95% B; ramp: 0.84-0.85 min 95% A, 5% B;0.85-1.5 min 95% A, 5% B.

MS/MS: WATERS Quattro Micro Tandem MS/MS; Z-Spray API interface; HPLC-MSinlet splitter 1:20; measurement in the ESI mode.

5.) In Vitro Clearance Determinations with Hepatocytes

Incubations with fresh primary hepatocytes are carried out at 37° C. ina total volume of 1.5 ml with a modified Janus® robot (Perkin Elmer)while shaking The incubations typically contain 1 million living livercells/ml, approx 1 μM substrate and 0.05 M potassium phosphate buffer(pH=7.4). The final acetonitrile concentration in the incubation is 1%.

Aliquots of 125 μl are withdrawn from the incubations after 2, 10, 20,30, 50, 70 and 90 min and transferred into 96-well filter plates (0.45μm low-binding hydrophilic PTFE; Millipore: MultiScreen Solvinert). Eachof these contain 250 μl of acetonitrile to stop the reaction. After thecentrifugation, the filtrates are analysed by MS/MS (typically API3000).

The in vitro clearances are calculated from the half-lives of thesubstance degradation, using the following equation:

CL′ _(intrinsic) [ml/(min·kg)]=(0.693/in vitro t1/2[min])·(liver weight[g liver/kg bodyweight])·(cell count[1.1·10̂8]/liver weight [g])/(cellcount[1·10̂6]/incubation volume [ml])

The CL_(blood) is calculated without taking into account the freefraction (“nonrestricted well stirred model”) by the following equation:

CL _(blood) well-stirred [l/(h·kg)]=(Q _(H) [l/(h·kg)]·CL′ _(intrinsic)[l/(h·kg)])/(Q _(H) [l/(h·kg)]+CL′_(intrinsic) [l/(h·kg)])

The species-specific extrapolation factors used for the calculation aresummarized in the following table:

male/female Mouse Mouse Rat Dog Cyno Man m f m/f m/f f m/f Cell number/g110 110 110 110 110 110 liver [10⁶ cells] Liver[g]/ 50 43 32 39 30 21 kgbodyweight Liver blood 5.4 5.4 4.2 2.1 2.5 1.3 flow [l/(h · kg)]

Fmax values which state the maximum possible bioavailability—based onthe hepatic extraction—are calculated as follows:

F _(max) well-stirred [%]=(1−(CL _(blood) well-stirred [l/(h·kg)]/Q _(H)[l/(h·kg)]))·100

6.) Determination of In Vivo Pharmacokinetics

To determine the in vivo pharmacokinetics, the test substances aredissolved in different formulation media (e.g. plasma, ethanol, DMSO,PEG400, etc.) or mixtures of these solubilizers, and administedintravenously or perorally to mice, rats, dogs or monkeys. Intravenousadministration is effected either as a bole or as an infusion. The dosesadministered are in the range from 0.1 to 5 mg/kg. Blood samples aretaken by means of a catheter or as sacrifice plasma at different timesover a period of up to 26 h. In addition, some organ, tissue and urinesamples are also obtained. The substances are determined quantitativelyin the test samples by means of calibration samples which areestablished in the particular matrix. Proteins present in the samplesare removed by precipitation with acetonitrile or methanol.Subsequently, the samples are separated by means of HPLC on a 2300 HTLCsystem (Cohesive Technologies, Franklin, Mass., USA) or Agilent 1200(Boblingen, Germany) using reverse-phase columns The HPLC system iscoupled via a turbo ion spray interface to an API 3000 or 4000 triplequadropole mass spectrometer (Applied Biosystems, Darmstadt, Germany).The plot of plasma concentration against time is evaluated using avalidated kinetics evaluation program.

C) WORKING EXAMPLES OF PHARMACEUTICAL COMPOSITIONS

The inventive substances can be converted in pharmaceutical preparationsas follows:

Tablet: Composition:

100 mg of the compound of Example 1, 50 mg of lactose (monohydrate), 50mg of maize starch, 10 mg of polyvinylpyrrolidone (PVP 25) (from BASF,Germany) and 2 mg of magnesium stearate.

Tablet weight 212 mg. Diameter 8 mm, radius of curvature 12 mm

Production:

The mixture of the compound of Example 1, lactose and starch isgranulated with a 5% solution (m/m) of the PVP in water. The granulesare dried and then mixed with the magnesium stearate for 5 min. Thismixture is compressed in a conventional tablet press (see above fortablet format).

Oral Suspension: Composition:

1000 mg of the compound of Example 1, 1000 mg of ethanol (96%), 400 mgof Rhodigel (xanthan gum) (from FMC, USA) and 99 g of water.

A single dose of 100 mg of the inventive compound corresponds to 10 mlof oral suspension.

Production:

The Rhodigel is suspended in ethanol, and the compound of Example 1 isadded to the suspension. The water is added while stirring. The mixtureis stirred for approx. 6 h until the Rhodigel has finished swelling.

Intravenously Administrable Solution: Composition:

1 mg of the compound of Example 1, 15 g of polyethylene glycol 400 and250 g of water for injections.

Production:

The compound of Example 1 is dissolved together with polyethylene glycol400 by stirring in the water. The solution is sterile-filtered (porediameter 0.22 μm) and dispensed under aseptic conditions intoheat-sterilized infusion bottles. The latter are closed with infusionstoppers and crimped caps.

1-14. (canceled)
 15. A method of prophylaxis of thromboembolic disordersin humans and animals comprising administering to the human or animal ananticoagulatory amount of a compound of formula (I):

in which R¹ is trifluoromethyl, 1,1-difluoroethyl, 2,2-difluoroethyl,2,2,2-trifluoroethyl, difluoromethoxy, trifluoromethoxy or ethyl, R² is2-hydroxyeth-1-yl, 2-methoxyeth-1-yl, 2-ethoxyeth-1-yl, cyclopropyl or1-methoxycycloprop-1-yl, R³ is a group of the formula

where * is the point of attachment to the carbonyl group, or a saltthereof.
 16. The method of claim 15, wherein in the compound of formula(I), R¹ is trifluoromethyl, 2,2,2-trifluoroethyl, trifluoromethoxy orethyl, R² is 2-methoxyeth-1-yl, cyclopropyl or 1-methoxycycloprop-1-yl,R³ is a group of the formula

where * is the point of attachment to the carbonyl group.
 17. The methodof claim 15, wherein in the compound of formula (I), R¹ istrifluoromethoxy. R² is 2-methoxyeth-1-yl or cyclopropyl, and R³ is agroup of the formula

where * is the point of attachment to the carbonyl group.
 18. The methodof claim 15, wherein in the compound of formula (I) R¹ istrifluoromethoxy, R² is cyclopropyl, and R³ is a group of the formula

where * is the point of attachment to the carbonyl group.
 19. The methodof claim 15, wherein the phenyl substituent and the 1,2,4-oxadiazol-5-ylsubstituent of the compound of formula (I) bonded to the piperidine ringare in cis positions to one another.
 20. The method of claim 15, whereinthe carbon atom to which the phenyl substituent is bonded in thecompound of formula (I) is in an S configuration and the carbon atom towhich the 1,2,4-oxadiazol-5-yl substituent is bonded is also in an Sconfiguration.
 21. The method of claim 15, wherein the compound offormula (I) is has the formula:


22. The method of claim 21, wherein the compound is a cis stereoisomer.23. The method of claim 21, wherein the compound of formula (I) is{3-(3-Cyclopropyl-1,2,4-oxadiazol-5-yl)-5-[4-(trifluoromethoxy)phenyl]piperidin-1-yl}(1,1-dioxidothiomorpholin-4-yl)methanone[cis isomer], or a salt thereof.
 24. The method of claim 15, wherein thecompound of formula (I) is administered as a pharmaceutical composition,comprising the compound of formula (I) and an inert nontoxicpharmaceutically acceptable excipient.
 25. The method of claim 15,wherein the thromboembolic disorder is selected from the groupconsisting of: ST-segment elevation myocardial infarction (STEMI) andnon-ST-segment elevation myocardial infarction (non-STEMI), stableangina pectoris, unstable angina pectoris, reocclusions and restenosesafter coronary interventions such as angioplasty, stent implantations oraortocoronary bypass, peripheral arterial occlusion diseases, pulmonaryembolisms, deep venous thromboses and renal vein thromboses, transitoryischaemic attacks and also thrombotic and thromboembolic stroke.
 26. Themethod of claim 15, wherein the condition is a cardiogenicthromboembolism selected from the group consisting of brain ischaemias,stroke and systemic thromboembolisms and ischaemias; an acute,intermittent or persistent cardiac arrhythmia
 27. The method of claim15, wherein the condition is a thromboembolic complication connectedwith microangiopathic haemolytic anaemias, extracorporeal circulation.28. A method of prophylaxis of a pulmonary disorder comprisingadministering to a patient in need thereof a therapeutically effective,anticoagulatory, amount of a compound of formula (I):

in which R¹ is trifluoromethyl, 1,1-difluoroethyl, 2,2-difluoroethyl,2,2,2-trifluoroethyl, difluoromethoxy, trifluoromethoxy or ethyl, R² is2-hydroxyeth-1-yl, 2-methoxyeth-1-yl, 2-ethoxyeth-1-yl, cyclopropyl or1-methoxycycloprop-1-yl, R³ is a group of the formula

where * is the point of attachment to the carbonyl group, or a saltthereof.
 29. The method of claim 28, wherein the compound of formula (I)is administered as a pharmaceutical composition, comprising the compoundof formula (I) and an inert nontoxic pharmaceutically acceptableexcipient.
 30. The method of claim 28, wherein the pulmonary disorder isselected from the group consisting of pulmonary fibrosis; pulmonaryhypertension, including pulmonary arterial hypertension, and disorderscharacterized by pulmonary occlusion; and an inflammatory pulmonarydisorder.